DDBJ/EMBL/GenBank accession nos AF038034-AF038042
Germline alterations of BRCA1 result in susceptibility to breast and ovarian cancer. The protein encoded by BRCA1 interacts in vivo with the BRCA1-associated RING domain (BARD1) protein. Accordingly, BARD1 is likely to be a critical factor in BRCA1-mediated tumor suppression and may also serve as a target for tumorigenic lesions in some human cancers. We have now determined the genomic structure of BARD1 and performed a mutational analysis of 58 ovarian tumors, 50 breast tumors and 60 uterine tumors. Seven polymorphisms were detected within the 2.34 kb coding sequence of BARD1. Somatically acquired missense mutations were observed in one breast carcinoma and one endometrial tumor; in at least one of these cases, tumor formation was accompanied by loss of the wild-type BARD1 allele, following the paradigm for known tumor suppressor genes. In addition, a germline alteration of BARD1 was identified in a clear cell ovarian tumor (Gln564His); again, loss of the wild-type BARD1 allele was observed in the malignant cells of this patient. The Gln564His patient was also diagnosed with two other primary cancers: a synchronous lobular breast carcinoma and a stage IA clear cell endometrioid cancer confined to an endometrial polyp 6 years earlier. These findings suggest an occasional role for BARD1 mutations in the development of sporadic and hereditary tumors.
Alterations in the BRCA1 gene (1,2) account for 40-50% of early onset familial breast cancer cases and >75% of cases of familial breast/ovarian cancer (3,4). In families with germline BRCA1 alterations, tumor formation requires loss or alteration of the wild-type allele (5), in keeping with the hypothesis that BRCA1 has tumor suppressor function.
BRCA1 encodes a large polypeptide of 1863 amino acids that contains an amino-terminal RING domain and two carboxy-terminal BRCT domains (2,6). Although the functions of these amino acid motifs are not precisely determined, the RING and BRCT domains of BRCA1 are well conserved phylogenetically and serve as common sites for missense mutations that predispose women to early-onset breast cancer. Therefore, these domains appear to be critical for tumor suppression by BRCA1.
We recently described an interaction between BRCA1 and a protein termed BARD1 (the BRCA1-associated RING-domain protein) (7). BARD1 is structurally related to BRCA1 in that it harbors an amino-terminal RING domain and two carboxy-terminal BRCT motifs. Studies of their subcellular distributions have shown that both proteins co-localize to discrete nuclear structures at defined stages of cell cycle progression (8,9). The in vivo association between BRCA1 and BARD1 is mediated by sequences within their respective RING domains. The fact that disease-related missense mutations within the RING motif of BRCA1 ablate the BRCA1-BARD1 interaction suggests that this protein complex mediates the tumor suppression function ascribed to BRCA1 (7).
If the BRCA1-BARD1 complex is indeed essential for tumor suppressor activity, then it is reasonable to ask if alterations of BARD1 are also involved in oncogenesis. Therefore, we determined the genomic structure of BARD1 and confirmed its localization by fluorescence in situ hybridization (FISH) to 2q34-q35, a region that is not frequently altered in breast or ovarian tumors. We also screened a panel of human tumors for genetic alterations of BARD1. Of these, 1/58 ovarian tumors, 1/50 breast tumors and 1/60 uterine tumors harbored different missense alterations, each of which lies within or close to the BRCT domains of BARD1. The wild-type allele of BARD1 was clearly absent in both the ovarian and uterine tumors, implying that loss of BARD1 function may contribute to oncogenesis. Although the BARD1 mutations in the breast and uterine tumors (Val695Leu and Ser761Asn) were acquired somatically, the ovarian tumor mutation (Gln564His) was of germline derivation. This patient had also developed independent primary cancers of the breast and endometrium. Thus, although further studies are needed, these data imply that loss of BARD1 function may be a contributing factor in the formation of some sporadic and hereditary tumors.
To obtain the genomic DNA encoding BARD1, bacteriophage [lambda] and BAC (bacterial artificial chromosome) libraries of human genomic DNA were screened by hybridization with fragments of BARD1 cDNA (7). Eleven hybridizing [lambda] clones and two hybridizing BAC clones were subjected to nucleotide sequence analysis with oligonucleotide primers derived from the BARD1 cDNA sequence. This analysis revealed that the BARD1-coding sequences are derived from 11 exons distributed over at least 65 kb of genomic DNA [GenBank accession nos AF038034 (exon I), AF038035 (exons II and III), AF038036 (exon IV), AF038037 (exon V), AF038038 (exon VI), AF038039 (exon VII), AF038040 (exon VIII), AF038041 (exon IX) and AF038042 (exon X)]. The chromosomal origin of BARD1 was then established by FISH of normal human metaphase chromosomes with plasmid subclones containing BARD1 genomic sequences. FISH analysis localized BARD1 to bands 2q34-35, consistent with the BARD1 mapping data obtained previously with the Genebridge panel of whole genome radiation hybrid DNAs (7).
The primer sets that were used to amplify the 11 coding exons of BARD1 from cDNA or genomic DNA templates are presented in Table 1.
Table 1.
Table 2
Table 3
We used single-stranded conformational polymorphism (SSCP) analysis (10) to screen genomic DNAs or cDNAs from 50 breast tumors, 58 ovarian tumors, 60 uterine cancers (primarily endometrial), six breast cancer lines and six ovarian cancer lines for genetic alterations in BARD1. Variant bands were excised from the SSCP gel, subjected to a second round of amplification and DNA sequenced.
The BARD1 polypeptide is comprised of 777 amino acids and includes an amino-terminal RING domain (residues 46-90), three tandem ankyrin repeats (residues 427-525) and two carboxy-terminal BRCT motifs (residues 605-777) (see Fig. 1). Seven polymorphisms were detected within the 2.34 kb of BARD1-coding sequence (Table 2 and Fig. 1). One of these is an in-frame deletion of seven amino acids between its RING domain and the ankyrin repeats (residues 358-364). This deletion was seen in 1% of Caucasian and 7% of unrelated African chromosomes. Given the low frequency of this allele in Caucasians, it was surprising to find that both the MCF7 breast cancer cell line and the PEO4 ovarian cancer cell line harbored this deletion since they were both derived from Caucasian patients (11,12). However, gene deletions do not necessarily account for disease or cancer susceptibility. For example, a polymorphic stop codon within the 3' end of the coding sequence of BRCA2 results in loss of the 93 most terminal amino acids (Lys3326ter) with as yet no described deleterious effect (13).
When 58 ovarian tumors were analyzed with SSCP, one (ov61) was found to harbor a missense mutation within BARD1 that resulted in a glutamine to histidine (CAG -> CAC; Gln564His) change. This alteration falls between the ankyrin repeats and the BRCT domains of BARD1 (Figs 1 and 2, Table 3). The patient with this tumor was a woman of African-American origin who was diagnosed at age 73 with a clear cell adenocarcinoma of the ovary (stage 3A) and a synchronous infiltrating lobular carcinoma of the breast. SSCP analysis of RT-PCR products indicated that only the mutant transcript was present in the ovarian tumor mRNA from this individual, indicating that the wild-type transcript was either expressed at undetectable levels or was completely absent (Fig. 2).
To examine the BARD1 status in ov61 further, RT-PCR products from the tumor were cloned and sequenced and the fraction of clones with wild-type versus mutant sequence was determined. When 10 subclones were sequenced, all were found to harbor the mutant allele, indicating that the mRNA derived from the wild-type allele was truly absent in the ovarian tumor. When corresponding germline DNA from benign uterine tissue was cloned and sequenced, 4/9 clones harbored the mutant allele and 5/9 clones harbored the wild-type allele, indicating that the alteration in the tumor was of germline origin.
At the time of hysterectomy 6 years earlier, this patient had been diagnosed with an incidental stage IA endometrial clear cell tumor. It is likely that the tumors of the endometrium and ovary were independent primaries since the initial endometrial tumor was a small focus of carcinoma confined to an endometrial polyp.
We used SSCP analysis to examine genomic DNA extracted from paraffin-embedded tissue obtained from the three primary tumors from this patient as well as from benign uterine tissue (Fig. 3). This revealed the variant allele in all samples, including normal uterine tissue, again confirming that this alteration was of germline origin. Moreover, the wild-type allele of BARD1 was reduced in the genomic DNA of the ovarian tumor, explaining the loss of wild-type BARD1 transcripts. We detected both the wild-type and mutant alleles in genomic DNA of both the endometrial and breast cancers; however, a large proportion of the paraffin sections was normal tissue, so that it was not possible to determine if the wild-type allele had been lost in these tumors. Further histological examination indicated that a significant proportion of normal tissue had infiltrated these tumor specimens rendering microdissection of tumor tissue from these samples impossible.
The diagnosis of endometrial cancer in the patient with the Gln564His mutation prompted us to examine an additional 60 uterine cancers for genetic alterations in BARD1. One additional endometrial tumor (ut8) was identified with a novel BARD1 alteration in a patient of European origin who had been diagnosed at age 50 with a stage 4B malignant mixed mullerian tumor (carcinosarcoma) (Table 3). This alteration, a Ser761Asn change in the second BRCT domain, was not seen in >600 other Caucasian chromosomes by allele-specific oligonucleotide (ASO)/dot-blot analysis. Direct sequencing of genomic DNA derived from this tumor only revealed the mutant allele, indicating that the wild-type allele had again been lost. Microdissection of normal DNA from archival sections of this tumor followed by direct sequencing revealed only a wild-type allele indicating that the alteration was somatic in origin.
Finally, a breast tumor (BC64) obtained from a 60-year-old woman, and diagnosed as an invasive ductal carcinoma, was found to harbor a BARD1 alteration (Val695Leu) in one of the BRCT domains. The Val695Leu substitution was not seen in the germline DNA of the patient (Table 3) or in >600 chromosomes, indicating that it arose somatically upon malignant transformation. It was not possible to determine whether the wild-type allele had been lost in the malignant cells of this patient since genomic DNA from microdissected tumor cells was not available.
The genomic structure of BARD1 and amplimers described here will enable mutational analysis of this gene in a variety of human tumors and cancer patients. FISH analysis confirms the localization of BARD1 to chromosome 2q, and sub-localizes it to within 2q34-35, a region not shown to be altered frequently in neoplasia.
Screening a panel of primary breast (n = 50), ovarian (n = 58) and uterine (n = 60) tumors revealed three distinct missense alterations in the BARD1-coding sequence. One of these was a clear cell ovarian tumor that harbored a Gln564His mutation of germline origin. Notably, the wild-type allele of BARD1 had been lost from the malignant ovarian cells, but not normal cells, of this patient. The patient was 73 years of age and had developed three independent primary tumors: the aforementioned clear cell ovarian carcinoma, a synchronous lobular breast carcinoma and, 6 years earlier, a clear cell endometrial cancer. Unfortunately, it was not possible to obtain a family history on this patient to determine if other carriers of Gln564His had also developed cancer. Insufficient material was available to rule out conclusively the presence of a germline BRCA1 alteration in this patient. The Gln564Arg alteration is of unknown significance, and it is conceivable that the loss of the wild-type allele in the ovarian tumor was driven by a mutation in a neighboring gene. However, it is of interest that this residue, along with flanking amino acids, is conserved in the mouse ortholog of BARD1 (NTGQR in both species; unpublished data).
The diagnosis of endometrial cancer in this patient 6 years earlier was intriguing and prompted us to investigate the frequency of BARD1 alterations in an additional 60 uterine tumors. This resulted in the identification of a somatic BARD1 alteration (Ser761Asn) in a malignant mixed mullerian tumor (Ut8). Again, the wild-type allele of BARD1 was clearly missing from this tumor. Moreover, the altered residue (Ser761) lies within the 30 amino acid core of the second BRCT domain, adjacent to an invariant tryptophan that is characteristic of the BRCT motif (6,14,15). Finally, a somatic alteration (Val695Leu) was detected in a breast tumor. This change also lies within the region containing the BARD1 BRCT domains.
The three alterations that we describe lie within, or close to, the two tandem BRCT domains of BARD1. While the functions of the BRCT motif are not known, the corresponding region of BRCA1 has been shown to have transactivational potential (16,17). BRCT domains are often found in proteins that function in the cell cycle checkpoints induced by DNA damage, including RAD9, XRCC1 and RAD4 (6,14,15). This, together with the findings that BRCA1 associates with RAD51 (18) and that BRCA1, BARD1 and RAD51 co-localize to nuclear dots in S phase but not in G1 (8,9), suggest that these proteins may be involved in cell cycle checkpoint control in response to DNA damage.
BARD1 is altered in only a small percentage of sporadic breast and ovarian tumors and, in this respect, is similar to BRCA1 and BRCA2. In the case of BRCA1, no genetic alterations have been detected in sporadic breast tumors; however, 10% of ovarian tumors harbor somatic BRCA1 mutations that result in protein truncations. In these tumors there is also loss of the wild type allele (19,20). In the case of BRCA2, four independent studies collectively identified two sporadic missense alterations and one truncating mutation in 281 primary breast cancers and two somatic alterations in 185 ovarian carcinomas (21-26).
The role of BARD1 in the development of uterine cancer is intriguing. BRCA1 alterations have not been described in this type of tumor, although some BRCA1 linked families occasionally have members with a linked BRCA1 mutant haplotype and endometrial cancer [e.g. family 4, (27)]. An analysis of patient ov61 suggests that germline mutations of BARD1 may confer a predisposition to certain neoplastic conditions and that studies of BARD1 mutations in familial cases of breast, ovarian, or uterine carcinoma may be warranted, particularly in those patients that develop synchronous or metachronous tumors of endometrial, breast or ovarian tissues.
Tumor tissue, matched normal tissue and blood specimens were obtained as part of protocols approved by the University of Texas Southwestern Medical Center Human Subjects Review Board, St Paul's Medical Center of Dallas, Medical City of Dallas and The Southern division of the Co-operative Human Tissue network. The breast cancers were primarily infiltrating ductal carcinomas. The ovarian carcinomas were of mixed histology although the majority were papillary serous carcinomas. Of the 60 uterine cancers, 44 were endometrial, 11 were mixed mullerian tumors and five were sarcomas. The following breast and ovarian cancer cell lines were obtained from the American Type Culture Collection: MCF-7, ZR75-1, BT-483, BT-20, T-47D, BT-474, OVCAR3, CAOV-3. BG-1 was kindly provided by Dr E.J. Modest, Boston University, and 2008 and 2774 have been described elsewhere (28,29). The ovarian cancer line PE04 was obtained from Dr Simon Langdon (Medical Oncology Unit, Western General Hospital, Edinburgh, Scotland). Tumors were frozen immediately in liquid nitrogen and stored at -70°C prior to RNA extraction. Buffy coat was prepared from blood. In some cases, DNA was prepared from paraffin-embedded tissue. RNA and cDNA were prepared by standard procedures.
A human genomic library cloned into bacteriophage [lambda] was first screened by hybridization with fragments of BARD1 cDNA (7). Eleven hybridizing [lambda] clones were identified and subjected to nucleotide sequence analysis with oligonucleotide primers derived from the BARD1 cDNA sequence.
This identified exon-intron boundaries for all exons except for VII, VIII and IX. To obtain genomic sequence of this region, three BACs were identified by hybridization with B230EX-1.3, a probe comprised of BARD1 cDNA residues 1229-2530 (accession no. U76638) (Research Genetics). DNA was prepared by standard methods. The same primers were used to sequence two of these BAC DNAs generating exon-intron boundary sequences for this region.
SSCP was performed as described elsewhere (10) with oligonucleotide primers for BARD1 with cDNA or genomic DNA as shown in Table 1. cDNA was derived from tumor, matched normal tissue or cell lines. Genomic DNA was obtained from tumor tissue, matched normal tissue, cell lines, blood and paraffin-embedded tissue.
PCR of tumor or blood DNA/cDNA was performed in 20 µl volumes containing 100 ng of cDNA or genomic DNA template; 1* PCR buffer (Perkin Elmer); 200 µM each dATP, dGTP, dCTP and dTTP; 10 pmol each primer (Gibco BRL); 0.3 µCi of [32P]dCTP (Amersham); and 0.5 U of Taq DNA polymerase (Perkin Elmer). PCR conditions were 30 cycles of 94°C, 30s; 55°C (or as specified for annealing temperatures in Table 1), 30s; 72°C, 30s. A final extension reaction at 72°C was performed for 1 min. Amplified samples were diluted 1:10 in formamide buffer (98% formamide, 10 mM EDTA, pH 8.0, 0.05% bromophenol blue, 0.05% xylene cyanol), denatured at 95°C for 5 min then cooled rapidly to 4°C. For each sample, 4 µl was loaded onto an SSCP gel and run at 8 W (constant power) for 8-16 h in 0.6* TBE at room temperature. Gels contained 0.5* MDE (AT Biochem), 0.6* TBE, 240 µl of 10% ammonium persulfate and 24 µl of TEMED. Duplicate gels were prepared with a supplement of 10% glycerol. Gels were subjected to autoradiography with or without being dried. Film was exposed for 12-24 h with an intensifying screen.
Variant bands were excised from the gel after alignment with the autoradiograph and purified with Qiaquick Gel Extraction kit (Cat #28706). DNA was resuspended in 20 µl of H2O and 5 µl was treated with 10 U of exonuclease I (manufacturer) and 2 U of shrimp alkaline phosphatase (manufacturer) at 37°C for 15 min. Following inactivation of this reaction with heat (80°C for 15 min), the DNA template was subjected to cycle sequencing with Thermosequenase (Amersham Life Science) and [[alpha]-33P]ddNTPs. Sequencing reactions were electrophoresed in 8% acrylamide/bis gels with 1* glycerol tolerant gel buffer at 70 W constant power for 2 h. Gels were dried and subjected to autoradiography.
Products obtained following PCR amplification were cloned directly into the TA-cloning vector pCR 2.1 (Invitrogen Cat #K2000-01). Cloning and transformation were performed according to the manufacturer's instructions. Colonies with inserts of the correct size were identified by direct amplification of the colony DNA in a PCR reaction with vector-specific primers (M13 forward/M13 reverse). In general, 70% of the colonies had inserts of the correct size; 30% of the colonies had no detectable insert. DNA was prepared from colonies with correct insert sizes according to standard procedures (30) and subjected to DNA sequencing with the M13 forward oligonucleotide primer and ABI DNA sequencing kit (Cat #402112). Sequencing reactions were analyzed with an ABI377.
PCR products were applied to replica Hybond filters as described elsewhere (31), except that instead of UV cross-linking, filters were baked for 2 h at 80°C.
Prehybridization and hybridization was performed as described elsewhere (32). The filters were subjected to a final wash in the Me4NCL solution for 2×20 min at 60°C.
To discriminate between the Ser761 and Asn761 alleles, PCR products were obtained with the US and UAS primers, and hybridized with the following ASOs: Asn761, GCTCCTTCGAACTGGTTTATA; Ser761, GCTCCTTCGAGCTGGTTTATA.
To obtain allele frequencies for the Arg658Cys polymorphism, PCR products obtained with R36H5/R36EAS primers were hybridized with: Cys658, TGAAGGTCCATGCAGAAGCAG; and Arg658, TGAAGGTCCACGCAGAAGCAG.
The cytogenetic location of BARD1 was obtained through FISH of normal human metaphase chromosome spreads with pooled DNA from three plasmid subclones in puc12 of [lambda] clones with inserts of 14, 16 and 13 kbp, representing 5', middle and 3' parts of the gene (R13S-14, R16S-16 and R36S-13) and containing a total of 43 kb of human sequence. Plasmid DNA was labeled with biotin by nick translation and subjected to FISH analysis as described elsewhere (33,34).
Drs Luca Cavalli-Sforza (Stanford University), Michele Ramsay and Trefor Jenkins (University of the Witwatersrand, South Africa) kindly provided DNA from Africans. Drs Wayne Taylor (Medical City of Dallas) and Jerry Shay (UT Southwestern Medical Center Tissue repository) provided breast tumor samples. Dr Michael Lovett provided thoughtful comments on the manuscript. We are indebted to Dr Vernie Stembridge for valuable support and advice. This work was supported in part by grants CA60650 (A.M.B) and CA76334 (R.B.) from the National Cancer Institute (NCI) and DE-FG03-96ER62173 (BT) from the US Department of energy. M.A.S. was supported in part by a grant from the Perot Family Foundation. Y.J. was supported in part by NCI training grant T32-CA09082.
Human Molecular Genetics Pages
Introduction
Results
Genomic structure of BARD1
BARD1 mutation screening
BARD1 polymorphisms
A germline BARD1 alteration
Somatic BARD1 alterations
Discussion
Materials And Methods
Clinical specimens
Genomic structure of BARD1
Mutational screening for BARD1 alterations
SSCP analysis
DNA sequencing of SSCP variants
Cloning of BARD1 PCR products for sequence analysis
PCR/ASO analysis of mutant alleles
FISH mapping of BARD1
Acknowledgements
References
Exon
Forward primer (5' -> 3')
Reverse primer (5' -> 3')
PCR product
size (bp)Template
Annealing
temp (°C)
I
R135S: CCGAGGAGCCTTTCATCCGA
R135AS: CGAGCGCGGCGCGACTGT
154
cDNA/genomic
59
I-III
B202-Z1S: ATGGAACCGGATGGTCGCGGT
B202-ZAS: TCTTCAAGTCTTGTATCCAGGC
213
cDNA
59
III-IV
B202-A: GCCTGGATACAAGACTTGAAG
B202-N:AACATCTGCAGGAGGACTTGG
306
cDNA
57
IV
B202-B: AAAGCTTCAGTGCAAACCCA
B202-BAS: TCCAGATCTTGCAGAAGCC
132
cDNA
53
IV
B202-X: CAGATGTTTCTGAGAGGGCT
B202-XAS: ATTCCTCTTTGGAGTCAAATTC
138
cDNA/genomic
55
IV
B230-A: GAGGCAGAAAAAGAAGATGGT
B230-AS: AGGAGCCACTTGCTAGTAAG
136
cDNA/genomic
55
IV
B202-Y: ATGGTGAAATAGACTTACTAGC
B202-YAS: GCAGACCTTCTCAGGAGTC
149
cDNA/genomic
55
IV
B230-B: AAGAGCAGGAATGAAGTAGTG
B230-BAS: CTCCACTGGTGCTCAGAATG
163
cDNA/genomic
55
IV
B230-C: AGTGGAGATTTTGTTAAGCAA
B230-CAS: AGGTGGTGTACCTGGTGAA
159
cDNA/genomic
51
IV
B230-D: GGTACACCACCTTCTACATT
B230-DAS: GTCTCTCCTCTATGATTTCTT
113
cDNA
53
IV-V
B230-PS: CAATGAAGCTGTTGCCCAA
B230-P: GTCTTTAACATTTGGATCACT
137
cDNA
51
V-VI
B230-E: AGTGATCCAAATGTTAAAGAC
B230-EAS: CCCATTCTTGGCTGCATC
162
cDNA
51
VI-VII
B230-F: CAAAATGACTCACCACTTCAC
B230-FAS: ATCGACAGGCCGCAGACC
120
cDNA
55
VII-IX
B230-FF: CCTGTCGATTATACAGATGAT
B230-FFAS: AACATGAGTTACTGTACTGTC
234
cDNA
57
VIII-X
B230-WS: TATACTGAGTTTGACAGTACAG
B230-WAS: CATACTTTTCTTCGTAGACATG
146
cDNA
55
X-XI
B230-G: TGGGTAAAAGCATGTCTACGA
B230-R: GTAGCATCCATCAAACAGCTT
126
cDNA
55
XI
B230-H: GGATGCTACTTCTATTTGTG
B230-HAS: GAGTCACGTCACTGTCTG
124
cDNA
51
XI
B230-TS: CCTCAGTAGAAAGCCCAAGC
B230-TAS: GCCCTGCCGAACCCTCTC
154
cDNA
57
XI
B230-US: GAGAGGGTTCGGCAGGGC
B230-UAS: TTCAATTTGAAATGTTCATCTGGT
124
cDNA/genomic
57
I
R1352S: ACAGTCGCGCCGCGCTCGA
R13AAS: CAGAAACTGTGCGACCCGTG
107
genomic
59
II
R12AS: AGATGTTTATCTAACAATGACTC
R12BAS: AGTTGTACTATATACATCAAACC
146
genomic
55
III
R13BS: ATTCTGCTGAATGGGTTGCTT
R13CAS: TAACTAAGAGAGATAGGGATAG
226
genomic
55
IV
R5C5: GGAGCTCCATGTGGGAGCAA
B202-N: AACATCTGCAGGAGGACTTGG
270
genomic
59
IV
B230-D: GGTACACCACCTTCTACATT
R5DAS: TCTGAGATGGTATTTCAGAGT
170
genomic
53
V
R34DS: TGCTTTTTAATTTCCATTTTGTTC
R34FAS: AAGAACTGTAAAACACAGAAAGA
163
genomic
55
VI
R34FS: TGCTCTTTCTTATCACTTCTTTC
R34GAS: CTTGACTCAAGAATATAGGTCC
278
genomic
57
VII
FFGS2: TTGAGTCGAGTCACACATTTGA
B2305FGAS: CTATTATGTTCCTTTCATAACCA
233
genomic
55
VIII
3FGR: TAATGTCTTTGTCTAGTCGTCTAA
WSGAS: GGTAGTTCTCCAAAAGGATCA
264
genomic
55
VIII
3FGR: TAATGTCTTTGTCTAGTCGTCTAA
3F: TTCTGAAGACAGCCCACTGC
123
genomic
55
IX
B230IXS: GAGTTATAAGAAGCAGGCCAA
B230IXAS: ATTTCTTAATTCTCTCAAATCCAA
199
genomic
55
X
R36H5: TAGTGCTCACTTGATACTTAGT
R36EAS: CATAATAAGAACAATGAAAGTTGT
187
genomic
55
XI
R36E5: TTGATCTGCCTTTAACAAATG
B230-TAS: GCCCTGCCGAACCCTCTC
296
genomic
57
Altered BARD1 nucleotide
Amino acid change
Population
Allele
Frequency
No. chromosomes screened
C143T
Pro24Ser
Caucasian
Pro
0.59
22
Ser
0.41
African
Pro
0.82
22
Ser
0.18
A530G
Lys153Glu
Lys
n.d.
n.d.
Glu
n.d.
C1121G
Silent (Thr351)
C
n.d.
n.d.
G
n.d.
1140-1160 (Deletion of CCATTGCCTG- AATGTTCTTCA)
1139del21 (del aa 358-364: PLPECSS)
Caucasian
WT
0.99
216
1139del21
0.01
African
WT
0.88
32
1139del21
0.12
A1592G
Val507Met
Met
n.d.
n.d.
Val
n.d.
G1765C
Cys557Ser
Caucasian
Cys
0.98
80
Ser
0.02
C2045T
Arg658Cys
Caucasian
Arg
0.98
138
Cys
0.02
African
Arg
1.00
42
Cys
0
Sample
(age)Tumor type
Codon
changeNucleotide
changeEffect
Loss of wild-
type alleleGermline
or somatic
ov61 (73 years)
Ovarian clear cell adenocarcinoma (Stage 3A)
CAG -> CAC
G1765C
Gln564His
Yes
Germline
ut8 (50 years)
Malignant mixed mulleriantumor (Stage 4B)
AGC -> AAC
G2354A
Ser761Asn
Yes
Somatic
BC64
Intraductal adenocarcinoma of the breast
GTC -> CTC
G2156C
Val695Leu
Nonedetectable
Somatic
REFERENCES
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 S-M Karppinen, R B Barkardottir, K Backenhorn, T Sydenham, K Syrjakoski, J Schleutker, T Ikonen, K Pylkas, K Rapakko, H Erkko, et al. Nordic collaborative study of the BARD1 Cys557Ser allele in 3956 patients with cancer: enrichment in familial BRCA1/BRCA2 mutation-negative breast cancer but not in other malignancies J. Med. Genet., November 1, 2006; 43(11): 856 - 862. [Abstract] [Full Text] [PDF]
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S. Sankaran, L. M. Starita, A. M. Simons, and J. D. Parvin Identification of Domains of BRCA1 Critical for the Ubiquitin-Dependent Inhibition of Centrosome Function. Cancer Res., April 15, 2006; 66(8): 4100 - 4107. [Abstract] [Full Text] [PDF]
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M K Sauer and I L Andrulis Identification and characterization of missense alterations in the BRCA1 associated RING domain (BARD1) gene in breast and ovarian cancer J. Med. Genet., August 1, 2005; 42(8): 633 - 638. [Abstract] [Full Text] [PDF]
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A. D. Choudhury, H. Xu, A. P. Modi, W. Zhang, T. Ludwig, and R. Baer Hyperphosphorylation of the BARD1 Tumor Suppressor in Mitotic Cells J. Biol. Chem., July 1, 2005; 280(26): 24669 - 24679. [Abstract] [Full Text] [PDF]
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S. Schuchner, V. Tembe, J. A. Rodriguez, and B. R. Henderson Nuclear Targeting and Cell Cycle Regulatory Function of Human BARD1 J. Biol. Chem., March 11, 2005; 280(10): 8855 - 8861. [Abstract] [Full Text] [PDF]
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 A. Feki, C.-E. Jefford, P. Durand, J. Harb, H. Lucas, K.-H. Krause, and I. Irminger-Finger BARD1 Expression During Spermatogenesis Is Associated with Apoptosis and Hormonally Regulated Biol Reprod, November 1, 2004; 71(5): 1614 - 1624. [Abstract] [Full Text] [PDF]
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S-M Karppinen, K Heikkinen, K Rapakko, and R Winqvist Mutation screening of the BARD1 gene: evidence for involvement of the Cys557Ser allele in hereditary susceptibility to breast cancer J. Med. Genet., September 1, 2004; 41(9): e114 - e114. [Full Text] [PDF]
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M. Rodriguez, X. Yu, J. Chen, and Z. Songyang Phosphopeptide Binding Specificities of BRCA1 COOH-terminal (BRCT) Domains J. Biol. Chem., December 26, 2003; 278(52): 52914 - 52918. [Abstract] [Full Text] [PDF]
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 U. K. Westermark, M. Reyngold, A. B. Olshen, R. Baer, M. Jasin, and M. E. Moynahan BARD1 Participates with BRCA1 in Homology-Directed Repair of Chromosome Breaks Mol. Cell. Biol., November 1, 2003; 23(21): 7926 - 7936. [Abstract] [Full Text] [PDF]
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 T I Orban and E Olah Emerging roles of BRCA1 alternative splicing Mol. Pathol., August 1, 2003; 56(4): 191 - 197. [Abstract] [Full Text] [PDF]
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E. E. McCarthy, J. T. Celebi, R. Baer, and T. Ludwig Loss of Bard1, the Heterodimeric Partner of the Brca1 Tumor Suppressor, Results in Early Embryonic Lethality and Chromosomal Instability Mol. Cell. Biol., July 15, 2003; 23(14): 5056 - 5063. [Abstract] [Full Text] [PDF]
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Y. Xia, G. M. Pao, H.-W. Chen, I. M. Verma, and T. Hunter Enhancement of BRCA1 E3 Ubiquitin Ligase Activity through Direct Interaction with the BARD1 Protein J. Biol. Chem., February 7, 2003; 278(7): 5255 - 5263. [Abstract] [Full Text] [PDF]
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M. Fabbro, J. A. Rodriguez, R. Baer, and B. R. Henderson BARD1 Induces BRCA1 Intranuclear Foci Formation by Increasing RING-dependent BRCA1 Nuclear Import and Inhibiting BRCA1 Nuclear Export J. Biol. Chem., June 7, 2002; 277(24): 21315 - 21324. [Abstract] [Full Text] [PDF]
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A. Chen, F. E. Kleiman, J. L. Manley, T. Ouchi, and Z.-Q. Pan Autoubiquitination of the BRCA1{middle dot}BARD1 RING Ubiquitin Ligase J. Biol. Chem., June 7, 2002; 277(24): 22085 - 22092. [Abstract] [Full Text] [PDF]
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J. R. Morris, N. H. Keep, and E. Solomon Identification of Residues Required for the Interaction of BARD1 with BRCA1 J. Biol. Chem., March 8, 2002; 277(11): 9382 - 9386. [Abstract] [Full Text] [PDF]
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F. Gautier, I. Irminger-Finger, M. Grégoire, K. Meflah, and J. Harb Identification of an Apoptotic Cleavage Product of BARD1 as an Autoantigen: A Potential Factor in the Antitumoral Response Mediated by Apoptotic Bodies Cancer Res., December 1, 2000; 60(24): 6895 - 6900. [Abstract] [Full Text]
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 D. H. Ly, D. J. Lockhart, R. A. Lerner, and P. G. Schultz Mitotic Misregulation and Human Aging Science, March 31, 2000; 287(5462): 2486 - 2492. [Abstract] [Full Text]
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W. Bruening, A. H. Prowse, D. C. Schultz, M. Holgado-Madruga, A. Wong, and A. K. Godwin Expression of OVCA1, a Candidate Tumor Suppressor, Is Reduced in Tumors and Inhibits Growth of Ovarian Cancer Cells Cancer Res., October 1, 1999; 59(19): 4973 - 4983. [Abstract] [Full Text] [PDF]
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F. E. Kleiman and J. L. Manley Functional Interaction of BRCA1-Associated BARD1 with Polyadenylation Factor CstF-50 Science, September 3, 1999; 285(5433): 1576 - 1579. [Abstract] [Full Text]
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 I. Irminger-Finger, J. V. Soriano, G. Vaudan, R. Montesano, and A.-P. Sappino In Vitro Repression of Brca1-associated RING Domain Gene, Bard1, Induces Phenotypic Changes in Mammary Epithelial Cells J. Cell Biol., November 30, 1998; 143(5): 1329 - 1339. [Abstract] [Full Text] [PDF]
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R. Hashizume, M. Fukuda, I. Maeda, H. Nishikawa, D. Oyake, Y. Yabuki, H. Ogata, and T. Ohta The RING Heterodimer BRCA1-BARD1 Is a Ubiquitin Ligase Inactivated by a Breast Cancer-derived Mutation J. Biol. Chem., April 27, 2001; 276(18): 14537 - 14540. [Abstract] [Full Text] [PDF]
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Y. Houvras, M. Benezra, H. Zhang, J. J. Manfredi, B. L. Weber, and J. D. Licht BRCA1 Physically and Functionally Interacts with ATF1 J. Biol. Chem., November 10, 2000; 275(46): 36230 - 36237. [Abstract] [Full Text] [PDF]
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V. Joukov, J. Chen, E. A. Fox, J. B. A. Green, and D. M. Livingston Functional communication between endogenous BRCA1 and its partner, BARD1, during Xenopus laevis development PNAS, October 9, 2001; 98(21): 12078 - 12083. [Abstract] [Full Text] [PDF]
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