In vivo biotinylation and capture of HIV-1 matrix and integrase proteins

Michael A. Belshan, Cameron J. Schweitzer, Meghan R. Donnellan, Richard Lu, Alan Engelman

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

This report describes the adaptation of the biotin ligase BirA-biotin acceptor sequence (BAS) labeling system to biotinylate specific human immunodeficiency virus 1 (HIV-1) proteins in vivo. Two HIV-1 clones were constructed, with the BAS introduced into the matrix region of gag or the integrase region of pol. Specific biotinylation of target proteins in virions was observed when molecular clones were co-expressed with BirA. Both BAS-containing viruses propagated in SupT1 T-cells although replication of the integrase clone was delayed. Further studies demonstrated that the integrase insertion yielded an approximate 40% reduction in single-round infectivity as assessed on MAGI-5 indicator cells, as well as in the in vitro integration activity of preintegration complexes extracted from acutely infected C8166-45 T-cells. Biotinylation of the integrase BAS tag furthermore rendered this virus non-infectious. The matrix viral clone by contrast displayed wild-type behavior under all conditions tested. These results therefore establish a system whereby biotinylated matrix protein in the context of replication-competent virus could be used to label and capture viral protein complexes in vivo.

Original languageEnglish
Pages (from-to)178-184
Number of pages7
JournalJournal of Virological Methods
Volume159
Issue number2
DOIs
StatePublished - Aug 2009

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Biotinylation
Integrases
Biotin
Clone Cells
HIV-1
Viruses
Human Immunodeficiency Virus Proteins
T-Lymphocytes
Viral Proteins
Ligases
Virus Replication
Virion
Proteins
p17 protein, Human Immunodeficiency Virus Type 1

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

In vivo biotinylation and capture of HIV-1 matrix and integrase proteins. / Belshan, Michael A.; Schweitzer, Cameron J.; Donnellan, Meghan R.; Lu, Richard; Engelman, Alan.

In: Journal of Virological Methods, Vol. 159, No. 2, 08.2009, p. 178-184.

Research output: Contribution to journalArticle

Belshan, Michael A. ; Schweitzer, Cameron J. ; Donnellan, Meghan R. ; Lu, Richard ; Engelman, Alan. / In vivo biotinylation and capture of HIV-1 matrix and integrase proteins. In: Journal of Virological Methods. 2009 ; Vol. 159, No. 2. pp. 178-184.
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