Isolation of the human PC6 gene encoding the putative host protease for HIV-1 gp160 processing in CD4+ T lymphocytes.
Production of infectious HIV-1 virions is dependent on the processing of envelope glycoprotein gp160 by a host cell protease. The protease in human CD4+ T lymphocytes has not been unequivocally identified, yet members of the family of mammalian subtilisin-like protein convertases (SPCs), which are soluble or membrane-bound proteases of the secretory pathway, best fulfill the criteria. These proteases are required for proprotein maturation and cleave at paired basic amino acid motifs in numerous cellular and viral glycoprotein precursors, both in vivo and in vitro. To identify the gp160 processing protease, we have used reverse transcription-PCR and Northern blot analyses to ascertain the spectrum of SPC proteases in human CD4+ T cells. We have cloned novel members of the SPC family, known as the human PC6 genes. Two isoforms of the hPC6 protease are expressed in human T cells, hPC6A and the larger hPC6B. The patterns of SPC gene expression in human T cells has been compared with the furin-defective LoVo cell line, both of which are competent in the production of infectious HIV virions. This comparison led to the conclusion that the hPC6 gene products are the most likely candidates for the host cell protease responsible for HIV-1 gp160 processing in human CD4+ T cells.
Miranda
L
L
Department of Cellular and Structural Biology, Molecular Biology Program, University of Colorado Cancer Center, Denver 80262, USA.
Wolf
J
J
Pichuantes
S
S
Duke
R
R
Franzusoff
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000201:enzymology* / Q000821:virology*; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D019302:HIV Envelope Protein gp160 / Q000096:biosynthesis*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D008322:Mammals; D008969:Molecular Sequence Data; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D045702:Proprotein Convertase 5; D012333:RNA, Messenger / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D012697:Serine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D019302:HIV Envelope Protein gp160; D012333:RNA, Messenger; D011994:Recombinant Proteins; D045702:Proprotein Convertase 5; D012697:Serine Endopeptidases