Structure of glycosylated and unglycosylated gag and gag-pol precursor proteins of Moloney murine leukemia virus.
Precursor polyproteins containing translational products of the gag gene of Moloney murine leukemia virus were purified by gel electrophoresis and cleaved into large fragments by hydroxylamine, mild acid hydrolysis, or cyanogen bromide. The hydroxylamine cleavage method (specific for asparagine-glycine bonds) was adapted especially for this study. The electrophoretic mobility and antigenicity of the fragments and, in some cases, the presence or absence of [35S]methionine revealed detailed information on the structure of Pr65gag, gPr80gag, and Pr75gag (the unglycosylated variant of gPr80gag formed in vivo in the presence of tunicamycin or in vitro in a reticulocyte cell-free system). When compared with Pr65gag, gPr80gag contains 7,000 daltons of additional amino acids, presumably as, or as part of, a leader sequence at or very close to its N terminus. We present evidence that this leader may have replaced part of the p15 sequence. Furthermore, gPr80gag contains three separate carbohydrate groups. One is attached to the presumed leader sequence or to the p15 domain, and two are attached to the p30 domain. Each of the Moloney murine leukemia virus gag precursor proteins Pr65gag, gPr80gag, and Pr75gag corresponds with a read-through product into the pol gene. We designated these products Pr180gag-pol, gPr200gag-pol, and Pr190gag-pol (the unglycosylated variant of gPr200gag-pol), respectively. gPr200gag-pol contains all of the extra amino acids and carbohydrate groups present in gPr80gag and at least one carbohydrate group in its pol sequences.
D000595:Amino Acid Sequence; D002241:Carbohydrates / Q000032:analysis; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D015683:Gene Products, gag; D006868:Hydrolysis; D019811:Hydroxylamine; D006898:Hydroxylamines; D008979:Moloney murine leukemia virus / Q000032:analysis*; D011498:Protein Precursors / Q000032:analysis* / Q000302:isolation & purification; D014415:Tunicamycin / Q000494:pharmacology; D014764:Viral Proteins / Q000032:analysis* / Q000302:isolation & purification
D002241:Carbohydrates; D015683:Gene Products, gag; D006898:Hydroxylamines; D011498:Protein Precursors; D014764:Viral Proteins; D014415:Tunicamycin; D019811:Hydroxylamine; D003488:Cyanogen Bromide