title: Human immunodeficiency virus type 1 employs the cellular dynein light chain 1 protein for reverse transcription through interaction with its integrase protein.
abstract: In this study, we examined the requirement for host dynein adapter proteins such as dynein light chain 1 (DYNLL1), dynein light chain Tctex-type 1 (DYNLT1), and p150(Glued) in early steps of human immunodeficiency virus type 1 (HIV-1) replication. We found that the knockdown (KD) of DYNLL1, but not DYNLT1 or p150(Glued), resulted in significantly lower levels of HIV-1 reverse transcription in cells. Following an attempt to determine how DYNLL1 could impact HIV-1 reverse transcription, we detected the DYNLL1 interaction with HIV-1 integrase (IN) but not with capsid (CA), matrix (MA), or reverse transcriptase (RT) protein. Furthermore, by mutational analysis of putative DYNLL1 interaction motifs in IN, we identified the motifs (52)GQVD and (250)VIQD in IN as essential for DYNLL1 interaction. The DYNLL1 interaction-defective IN mutant HIV-1 (HIV-1IN(Q53A/Q252A)) exhibited impaired reverse transcription. Through further investigations, we have also detected relatively smaller amounts of particulate CA in DYNLL1-KD cells or in infections with HIV-1IN(Q53A/Q252A) mutant virus. Overall, our study demonstrates the novel interaction between HIV-1 IN and cellular DYNLL1 proteins and suggests the requirement of this virus-cell interaction for proper uncoating and efficient reverse transcription of HIV-1.
  
  
  OBJECTIVE
  Host cellular DYNLL1, DYNLT1, and p150(Glued) proteins have been implicated in the replication of several viruses. However, their roles in HIV-1 replication have not been investigated. For the first time, we demonstrated that during viral infection, HIV-1 IN interacts with DYNLL1, and their interaction was found to have a role in proper uncoating and efficient reverse transcription of HIV-1. Thus, interaction of IN and DYNLL1 may be a potential target for future anti-HIV therapy. Moreover, while our study has evaluated the involvement of IN in HIV-1 uncoating and reverse transcription, it also predicts a possible mechanism by which IN contributes to these early viral replication steps.
authors.affiliation: Laboratory of Molecular Human Retrovirology, Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
authors.forename: Kallesh Danappa
authors.initials: KD
authors.lastname: Jayappa
authors.affiliation: Laboratory of Molecular Human Retrovirology, Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
authors.forename: Zhujun
authors.initials: Z
authors.lastname: Ao
authors.affiliation: Laboratory of Molecular Human Retrovirology, Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
authors.forename: Xiaoxia
authors.initials: X
authors.lastname: Wang
authors.affiliation: HIV-1 RNA Trafficking Laboratory, Lady Davis Institute at the Jewish General Hospital and McGill University, Montréal, Québec, Canada.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Mouland
authors.affiliation: Department of Immunology, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
authors.forename: Sudhanshu
authors.initials: S
authors.lastname: Shekhar
authors.affiliation: Department of Immunology, Faculty of Medicine, University of Manitoba, Winnipeg, Canada.
authors.forename: Xi
authors.initials: X
authors.lastname: Yang
authors.affiliation: Laboratory of Molecular Human Retrovirology, Department of Medical Microbiology, Faculty of Medicine, University of Manitoba, Winnipeg, Canada yao2@cc.umanitoba.ca.
authors.forename: Xiaojian
authors.initials: X
authors.lastname: Yao
mesh_terms: D020816:Amino Acid Motifs; D002460:Cell Line; D056445:Cytoplasmic Dyneins / Q000378:metabolism*; D004252:DNA Mutational Analysis; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism; D055785:Gene Knockdown Techniques; D019427:HIV Integrase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D048348:Reverse Transcription; D057646:Virus Uncoating
chemical_list: C000606360:DCTN1 protein, human; C533376:DYNLT1 protein, human; D000072159:Dynactin Complex; D008869:Microtubule-Associated Proteins; D019427:HIV Integrase; C427758:DYNLL1 protein, human; D056445:Cytoplasmic Dyneins; D004398:Dyneins