PMID- 25491356
OWN - NLM
STAT- MEDLINE
DCOM- 20150626
LR  - 20231019
IS  - 2150-7511 (Electronic)
VI  - 5
IP  - 6
DP  - 2014 Dec 9
TI  - Membrane binding and subcellular localization of retroviral Gag proteins are 
      differentially regulated by MA interactions with 
      phosphatidylinositol-(4,5)-bisphosphate and RNA.
PG  - e02202
LID - 10.1128/mBio.02202-14 [doi]
LID - e02202-14
AB  - The matrix (MA) domain of HIV-1 mediates proper Gag localization and membrane 
      binding via interaction with a plasma-membrane (PM)-specific acidic phospholipid, 
      phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2]. HIV-1 MA also interacts with 
      RNA, which prevents Gag from binding to membranes containing phosphatidylserine, 
      a prevalent cellular acidic phospholipid. These results suggest that the MA-bound 
      RNA promotes PM-specific localization of HIV-1 Gag by blocking nonspecific 
      interactions with cellular membranes that do not contain PI(4,5)P2. To examine 
      whether PI(4,5)P2 dependence and RNA-mediated inhibition collectively determine 
      MA phenotypes across a broad range of retroviruses and elucidate the significance 
      of their interrelationships, we compared a panel of Gag-leucine zipper constructs 
      (GagLZ) containing MA of different retroviruses. We found that in vitro membrane 
      binding of GagLZ via HIV-1 MA and Rous sarcoma virus (RSV) MA is both PI(4,5)P2 
      dependent and susceptible to RNA-mediated inhibition. The PM-specific 
      localization and virus-like particle (VLP) release of these GagLZ proteins are 
      severely impaired by overexpression of a PI(4,5)P2-depleting enzyme, 
      polyphosphoinositide 5-phosphatase IV (5ptaseIV). In contrast, membrane binding 
      of GagLZ constructs that contain human T-lymphotropic virus type 1 (HTLV-1) MA, 
      murine leukemia virus (MLV) MA, and human endogenous retrovirus K (HERV-K) MA is 
      PI(4,5)P2 independent and not blocked by RNA. The PM localization and VLP release 
      of these GagLZ chimeras were much less sensitive to 5ptaseIV expression. Notably, 
      single amino acid substitutions that confer a large basic patch rendered HTLV-1 
      MA susceptible to the RNA-mediated block, suggesting that RNA readily blocks MA 
      containing a large basic patch, such as HIV-1 and RSV MA. Further analyses of 
      these MA mutants suggest a possibility that HIV-1 and RSV MA acquired PI(4,5)P2 
      dependence to alleviate the membrane binding block imposed by RNA. IMPORTANCE: MA 
      basic residues in the HIV-1 structural protein Gag interact with 
      phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P2] and RNA. RNA inhibits HIV-1 
      MA binding to non-PI(4,5)P2 acidic lipids. This inhibition may promote PM 
      specificity of Gag membrane binding, an early essential step in virus assembly. 
      However, whether and how relationships between these interactions have developed 
      among retroviruses are poorly understood. In this study, by comparing diverse 
      retroviral MA domains, we elucidated a strong correlation among PI(4,5)P2 
      dependence, susceptibility to RNA-mediated inhibition, and cellular behaviors of 
      Gag. Mutagenesis analyses suggest that a large basic patch on MA is sufficient to 
      confer susceptibility to RNA-mediated inhibition but not for PI(4,5)P2-dependent 
      membrane binding. Our findings highlight RNA's role as a general blocker of large 
      basic patches and suggest a possibility that some retroviruses, including HIV-1, 
      have evolved to bind PI(4,5)P2, while others have adopted smaller basic patches 
      on their MA domains, to overcome the RNA-mediated restriction of membrane 
      binding.
CI  - Copyright (c) 2014 Inlora et al.
FAU - Inlora, Jingga
AU  - Inlora J
AD  - Department of Microbiology and Immunology, University of Michigan Medical School, 
      Ann Arbor, Michigan, USA.
FAU - Collins, David R
AU  - Collins DR
AD  - Department of Microbiology and Immunology, University of Michigan Medical School, 
      Ann Arbor, Michigan, USA.
FAU - Trubin, Marc E
AU  - Trubin ME
AD  - Department of Microbiology and Immunology, University of Michigan Medical School, 
      Ann Arbor, Michigan, USA.
FAU - Chung, Ji Yeon J
AU  - Chung JY
AD  - Department of Microbiology and Immunology, University of Michigan Medical School, 
      Ann Arbor, Michigan, USA.
FAU - Ono, Akira
AU  - Ono A
AD  - Department of Microbiology and Immunology, University of Michigan Medical School, 
      Ann Arbor, Michigan, USA akiraono@umich.edu.
LA  - eng
GR  - R01 AI071727/AI/NIAID NIH HHS/United States
GR  - T32 AI007528/AI/NIAID NIH HHS/United States
GR  - T32 GM008353/GM/NIGMS NIH HHS/United States
GR  - T32 GM145304/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20141209
PL  - United States
TA  - mBio
JT  - mBio
JID - 101519231
RN  - 0 (Gene Products, gag)
RN  - 0 (Phosphatidylinositol 4,5-Diphosphate)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Cell Membrane/*virology
MH  - Gene Products, gag/*metabolism
MH  - HeLa Cells
MH  - Humans
MH  - Phosphatidylinositol 4,5-Diphosphate/*metabolism
MH  - Protein Binding
MH  - RNA, Viral/*metabolism
MH  - Retroviridae/*physiology
PMC - PMC4324246
EDAT- 2014/12/11 06:00
MHDA- 2015/06/27 06:00
PMCR- 2014/12/09
CRDT- 2014/12/11 06:00
PHST- 2014/12/11 06:00 [entrez]
PHST- 2014/12/11 06:00 [pubmed]
PHST- 2015/06/27 06:00 [medline]
PHST- 2014/12/09 00:00 [pmc-release]
AID - mBio.02202-14 [pii]
AID - mBio02202-14 [pii]
AID - 10.1128/mBio.02202-14 [doi]
PST - epublish
SO  - mBio. 2014 Dec 9;5(6):e02202. doi: 10.1128/mBio.02202-14.