Element-specific localization of Drosophila retrotransposon Gag proteins occurs in both nucleus and cytoplasm.
Many Drosophila non-long terminal repeat (LTR) retrotransposons actively transpose into internal, gene-rich regions of chromosomes but do not transpose onto chromosome ends. HeT-A and TART are remarkable exceptions; they form telomeres of Drosophila by repeated transpositions onto the ends of chromosomes and never transpose to internal regions of chromosomes. Both telomeric and nontelomeric, non-LTR elements transpose by target-primed reverse transcription, and their targets are not determined simply by DNA sequence, so it is not clear why these two kinds of elements have nonoverlapping transposition patterns. To explore roles of retrotransposon-encoded proteins in transposition, we analyzed intracellular targeting of Gag proteins from five non-LTR retrotransposons, HeT-A, TART, jockey, Doc, and I factor. All were expressed as green fluorescent protein-tagged proteins in cultured Drosophila cells. These Gag proteins have high levels of sequence similarity, but they have dramatic differences in intracellular targeting. As expected, HeT-A and TART Gags are transported efficiently to nuclei, where they show specific patterns of localization. These patterns are cell cycle-dependent, disappearing during mitosis. In contrast, only a fraction of jockey Gag moves into nuclei, whereas neither Doc nor I factor Gag is detected in the nucleus. Gags of the nontelomeric retrotransposons form characteristic clusters in the cytoplasm. These experiments demonstrate that closely related retrotransposon Gag proteins can have different intracellular localizations, presumably because they interact differently with cellular components. We suggest that these interactions reflect mechanisms by which the cell influences the level of transposition of an element.
Rashkova
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Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Karam
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Pardue
M-L
ML
D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D004330:Drosophila / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics*; D018626:Retroelements / Q000235:genetics*; D020079:Terminal Repeat Sequences / Q000235:genetics; D013997:Time Factors
D029721:Drosophila Proteins; D011993:Recombinant Fusion Proteins; D018626:Retroelements