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Specific HIV‐1 integrase‐reverse transcriptase interactions as a mechanism for promoting reverse transcription
Author(s) -
Chow Samson A.,
Wilkinson Thomas A,
Zhang Min,
Januszyk Kurt,
Phillips Martin L,
Le Grice Stuart F. J.,
Clubb Robert T
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.501.1
Subject(s) - reverse transcriptase , integrase , transcription (linguistics) , biology , viral replication , microbiology and biotechnology , chemistry , virus , virology , human immunodeficiency virus (hiv) , gene , genetics , rna , linguistics , philosophy
Integrase (IN) from human immunodeficiency virus‐type 1 (HIV‐1) exerts pleiotropic effects in the viral replication cycle, and IN mutations can impair reverse transcription. We showed previously that IN stimulates both the initiation and elongation modes of reverse transcription by increasing the processivity of reverse transcriptase (RT) and suppressing the formation of the pause products. IN and RT interact in vitro, and the IN C‐terminal domain (CTD) is both necessary and sufficient for binding RT. We used nuclear magnetic resonance spectroscopy to identify a putative RT‐binding surface on the IN CTD, and surface plasmon resonance to obtain kinetic parameters and the binding affinity for the IN‐RT interaction. An IN K258A substitution that disrupts reverse transcription in infected cells is located at the putative RT‐binding surface, and we find that this substitution substantially weakens IN CTD‐RT interactions. Other substitutions at the putative RT‐binding surface also rendered the virus replication‐incompetent. These results strengthen the notion that productive IN‐RT interactions are important for efficient reverse transcription during HIV‐1 replication, and provide insights into this interaction at the molecular level. This research is supported by NIH R21 AI077386.