Knockdown of MAP4 and DNAL1 produces a post-fusion and pre-nuclear translocation impairment in HIV-1 replication.
DNAL1 and MAP4 are both microtubule-associated proteins. These proteins were identified as HIV-1 dependency factors in a screen with wild-type HIV-1. In this study we demonstrate that knockdown using DNAL1 and MAP4 siRNAs and shRNAs inhibits HIV-1 infection regardless of envelope. Using a fusion assay, we show that DNAL1 and MAP4 do not impact fusion. By assaying for late reverse transcripts and 2-LTR circles, we show that DNAL1 and MAP4 inhibit both by approximately 50%. These results demonstrate that DNAL1 and MAP4 impact reverse transcription but not nuclear translocation. DNAL1 and MAP4 knockdown cells do not display cytoskeletal defects. Together these experiments indicate that DNAL1 and MAP4 may exert their functions in the HIV life cycle at reverse transcription, prior to nuclear translocation.
Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Lurie 9-290, Chicago, IL 60611, USA. d-gallo@northwestern.edu
Daniel E
DE
Gallo
Thomas J
TJ
Hope
D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000821:virology; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism / Q000821:virology; D015658:HIV Infections / Q000235:genetics; D016325:HIV Long Terminal Repeat / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D021381:Protein Transport; D034622:RNA Interference; D034741:RNA, Small Interfering; D012367:RNA, Viral; D048348:Reverse Transcription; D053586:Virus Internalization; D014779:Virus Replication
C569367:DNAL1 protein, human; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; D012367:RNA, Viral