Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 13-21 |
| Number of pages | 9 |
| Journal | Virology |
| Volume | 422 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 5 2012 |
Funding
We would like to thank the Northwestern University Flow Cytometry Core Facility for providing assistance with flow cytometry experiments. Fb-Luc, and pCG-Gag-Pol plasmids were kind gifts from Dr. Alan Engleman at the Dana Farber Cancer Institute. The N74D NL4-3-Luciferase plasmid was kindly provided by Dr. Vineet N. KewalRamani at NCI Frederick National Cancer Institute. We would also like to thank Dr. Greg Smith at Northwestern University Feinberg School of Medicine for providing mRFP-VP26 labeled HSV-1. This study was supported by the NIH R01 AI047770 (with associated diversity supplement), P50 GM082545 , and the James B Pendleton Charitable Trust .
Keywords
- Cytoskeleton
- DNAL1
- HIV
- MAP4
- Microtubules
- RTC
- Reverse transcription
ASJC Scopus subject areas
- Virology
