The human immunodeficiency virus type 1 carboxyl-terminal third of capsid sequence in Gag-Pol is essential but not sufficient for efficient incorporation of Pr160(gag-pol) into virus particles.
To elucidate the role of the C-terminal portion of Gag in the incorporation of human immunodeficiency virus type 1 (HIV-1) Gag-Pol into virus particles, a series of HIV-1 Gag-Pol mutants with deletions in the C-terminal gag sequence was constructed and viral incorporation of the Gag-Pol deletion mutants was analyzed using co-transfecting 293T cells with a Pr55(gag) expression plasmid. The biological function of the incorporated HIV-1 pol gene product was tested using an infectivity assay of the released virus particles which were pseudotyped with the murine leukemia virus Env. Analysis indicated that Gag-Pol deletion mutants, with a removal of the matrix (MA) and/or nucleocapsid (NC) or of the N-terminal two thirds of the gag coding sequence, could be incorporated efficiently into virus particles and produce significant amounts of infectious virions when assayed in a single-cycle infection assay. In contrast, mutations involving a deletion of the major homology region and the adjacent C-terminal capsid sequence significantly affected Gag-Pol incorporation. However, incorporation into virus particles of a Gag-Pol deletion mutant retaining both the major homology region and the adjacent C-terminal capsid intact was still severely impaired. This suggests that the capsid major homology region and the adjacent C-terminal capsid sequence in Gag-Pol are necessary but not sufficient for the incorporation of HIV-1 Pr160(gag-pol) into virus particles.
Chiu
Hsu-Chen
HC
Institute of Clinical Medicine, National Yang-Ming University School of Medicine and Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan.
Liao
Wei-Hao
WH
Chen
Szu-Wen
SW
Wang
Chin-Tien
CT
D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D015685:Fusion Proteins, gag-pol / Q000378:metabolism / Q000502:physiology*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D014771:Virion / Q000378:metabolism*
D017931:DNA Primers; D015685:Fusion Proteins, gag-pol