Document Detail


Proteomic analysis of early HIV-1 nucleoprotein complexes.
MedLine Citation:
PMID:  23282062     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
After entry into the cell, the early steps of the human immunodeficiency virus type 1 (HIV-1) replication cycle are mediated by two functionally distinct nucleoprotein complexes, the reverse transcription complex (RTC) and preintegration complex (PIC). These two unique viral complexes are responsible for the conversion of the single-stranded RNA genome into double-stranded DNA, transport of the DNA into the nucleus, and integration of the viral DNA into the host cell chromosome. Prior biochemical analyses suggest that these complexes are large and contain multiple undiscovered host cell factors. In this study, functional HIV-1 RTCs and PICs were partially purified by velocity gradient centrifugation and fractionation, concentrated, trypsin digested, and analyzed by LC-MS/MS. A total of seven parallel infected and control biological replicates were completed. Database searches were performed with Proteome Discoverer and a comparison of the HIV-1 samples to parallel uninfected control samples was used to identify unique cellular factors. The analysis produced a total data set of 11055 proteins. Several previously characterized HIV-1 factors were identified, including XRCC6, TFRC, and HSP70. The presence of XRCC6 was confirmed in infected fractions and shown to be associated with HIV-1 DNA by immunoprecipitation-PCR experiments. Overall, the analysis identified 94 proteins unique in the infected fractions and 121 proteins unique to the control fractions with ≥ 2 protein assignments. An additional 54 and 52 were classified as enriched in the infected and control samples, respectively, based on a 3-fold difference in total Proteome Discoverer probability score. The differential expression of several candidate proteins was validated by Western blot analysis. This study contributes additional novel candidate proteins to the growing published bioinformatic data sets of proteins that contribute to HIV-1 replication.
Authors:
Cameron J Schweitzer; Teena Jagadish; Nicole Haverland; Pawel Ciborowski; Michael Belshan
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2013-01-16
Journal Detail:
Title:  Journal of proteome research     Volume:  12     ISSN:  1535-3907     ISO Abbreviation:  J. Proteome Res.     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-02-01     Completed Date:  2013-07-05     Revised Date:  2014-02-04    
Medline Journal Info:
Nlm Unique ID:  101128775     Medline TA:  J Proteome Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  559-72     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Antigens, CD / genetics,  metabolism
Antigens, Nuclear / genetics,  metabolism
Cell Line
Cell Nucleus / genetics,  metabolism,  virology*
Centrifugation, Density Gradient
Chromatography, Liquid
DNA, Viral / genetics*,  metabolism
DNA-Binding Proteins / genetics,  metabolism
Gene Expression Profiling
HIV-1 / genetics*,  metabolism
HSP70 Heat-Shock Proteins / genetics,  metabolism
Host-Pathogen Interactions
Humans
Lymphocytes / metabolism,  virology*
Nucleoproteins / genetics*,  metabolism
Protein Binding
Proteome / genetics*,  metabolism
Receptors, Transferrin / genetics,  metabolism
Reverse Transcription
Tandem Mass Spectrometry
Viral Proteins / genetics*,  metabolism
Virus Integration
Grant Support
ID/Acronym/Agency:
P30 MH062261/MH/NIMH NIH HHS; R01 AI080348/AI/NIAID NIH HHS; R01 DA030962/DA/NIDA NIH HHS; R01A1080348//PHS HHS; R01DA030962/DA/NIDA NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, CD; 0/Antigens, Nuclear; 0/CD71 antigen; 0/DNA, Viral; 0/DNA-Binding Proteins; 0/HSP70 Heat-Shock Proteins; 0/Ku autoantigen; 0/Nucleoproteins; 0/Proteome; 0/Receptors, Transferrin; 0/Viral Proteins
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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