A novel protease processing site in the transframe protein of human T-cell leukemia virus type 1 PR76(gag-pro) defines the N terminus of RT.
The genomic RNA of human T-cell leukemia virus type 1 encodes three polyproteins, Gag, Gag-Pro, and Gag-Pro-Pol, which are translated as a result of no, one, and two frameshifts, respectively. In this report we demonstrate that the 77 residues encoded at the C terminus of the Gag-Pro precursor can be collectively detected as an 8-kDa transframe protein (TFP) in virions. Mutant viruses with a C-terminally truncated TFP (19, 32, or 50 residues) had essentially a wild-type phenotype in vitro. However, a virus mutant that encoded only the Gag and Gag-Pro-Pol polyproteins due to a mutation in the second frameshift site, and hence did not produce TFP, was noninfectious. Mutation analysis of the proteolytic cleavage site between PR and TFP revealed the presence of an additional site and the existence of a p1 peptide separating protease and TFP. While removal of the cleavage site at the PR-p1 junction had a modest effect on virus replication, mutation of the p1-TFP cleavage site led to noninfectious virus and the loss of reverse transcriptase activity. Determination of the amino-terminal sequence of a hemagglutinin-tagged RT demonstrated that the same site is used in processing the Gag-Pro-Pol precursor and thus defines the start of mature RT. Neither mutation alone or in combination caused changes in the amounts or processing patterns of the Gag polyprotein, indicating that protease is active independent of its C terminus.
Heidecker
Gisela
G
Basic Research Laboratory, NCI-Frederick, Building 567, Room 154, Frederick, MD 21702-1201, USA. heidecke@ncifcrf.gov
Hill
Shawn
S
Lloyd
Patricia A
PA
Derse
David
D
D000595:Amino Acid Sequence; D016282:Aspartic Acid Endopeptidases / Q000378:metabolism*; D001483:Base Sequence; D015683:Gene Products, gag / Q000737:chemistry*; D015368:Human T-lymphotropic virus 1 / Q000737:chemistry*; D008969:Molecular Sequence Data; D009154:Mutation; D011498:Protein Precursors / Q000737:chemistry*; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry*
D015683:Gene Products, gag; D011498:Protein Precursors; D012194:RNA-Directed DNA Polymerase; D016282:Aspartic Acid Endopeptidases; C079711:HTLV-1 protease