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Human Molecular Genetics Advance Access published online on March 17, 2004
Human Molecular Genetics, doi:10.1093/hmg/ddh118
© 2004 by Oxford University Press
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©2004 Oxford University Press

Article

A Potential Role for the Nucleolus in L1 Retrotransposition

John L. Goodier 1*, Eric M. Ostertag 2, Kurt A. Engleka 3, Maria C. Seleme 3, and Haig H. Kazazian Jr.3

1 Department of Genetics, University of Pennsylvania School of Medicine, Rm. 515 CRB, 415 Curie Blvd., Philadelphia, PA 19104
2 Department of Genetics, University of Pennsylvania School of Medicine, Philadelphia; Department of Pathology and Laboratory Medicine, Hospital of the University of Pennsylvania, Philadelphia
3 Department of Genetics, University of Pennsylvania School of Medicine, Philadelphia

* To whom correspondence should be addressed. E-mail: jgoodier{at}mail.med.upenn.edu.


  Abstract

Determining the subcellular localization of the L1 ORF2 protein (ORF2p) has been impossible to date because of technical limitations in detecting either endogenous or overexpressed forms of the protein. Here we report visualization of the full-length ORF2p in cultured human cells following expression in a modified vaccinia virus/T7 RNA polymerase (MVA/T7RP) system. The MVA/T7RP system was used to ascertain subcellular localization of L1 ORF1p and ORF2p both as fusions with green fluorescent protein and by immunocytochemistry. Full-length ORF2p was predominantly cytoplasmic, while carboxy-terminal-deleted ORF2p localized additionally to the nucleolus. We mapped a functional nucleolar localization signal in ORF2p. ORF1p appeared in the cytoplasm with a speckled pattern and colocalized with ORF2p in nucleoli in a subset of cells. These findings help explain the presence of chimeras between L1s and small RNA gene sequences recently discovered in the human genome.


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