The human cytomegalovirus 86-kilodalton immediate-early 2 protein: synthesis as a precursor polypeptide and interaction with a 75-kilodalton protein of probable viral origin.
The immediate-early 2 (IE2) 86-kDa polypeptide, a major immediate-early gene product of human cytomegalovirus, regulates transcription both positively and negatively. We report two new properties of the IE2 86-kDa polypeptide in infected cells. Immunoprecipitation of infected cell proteins from human embryonic lung cells by antipeptide or monoclonal antibodies specific for IE2 epitopes revealed three closely migrating polypeptide species. The slowest, p86, behaved as expected for the mature 86-kDa IE2 polypeptide. The middle species, p80, was immunoprecipitated from denatured as well as native samples and labeled to steady state rapidly. Pulse-chase analysis demonstrated directly that p80 was a metabolic precursor to p86. The fastest-migrating species, p75, was not detected by probing blots of the immunoprecipitated proteins with IE2-specific antisera; p75 was not precipitated from denatured protein samples; and the products of partial proteolysis of p75 were distinct from those of p86. These properties established p75 as an unrelated coprecipitated polypeptide complexed with p86. The p75 proteins coprecipitated from cells infected with two different strains of human cytomegalovirus, AD169 and Towne, had different mobilities. p75 was detected as early as 6 h and as late as 72 h after infection, but it was not synthesized in cells released from a cycloheximide block. Thus, it is likely that p75 is an early viral protein.
Samaniego
L A
LA
Department of Microbiology and Immunology, Pennsylvania State University, College of Medicine, Hershey 17033.
Tevethia
M J
MJ
Spector
D J
DJ
D000914:Antibodies, Viral; D000918:Antibody Specificity; D002478:Cells, Cultured; D003429:Cross Reactions; D003587:Cytomegalovirus / Q000378:metabolism*; D006801:Humans; D017874:Immediate-Early Proteins / Q000096:biosynthesis* / Q000378:metabolism*; D008562:Membrane Glycoproteins; D010449:Peptide Mapping; D011233:Precipitin Tests; D011485:Protein Binding; D011498:Protein Precursors / Q000378:metabolism*; D015534:Trans-Activators; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins / Q000096:biosynthesis*; D014764:Viral Proteins / Q000378:metabolism
D000914:Antibodies, Viral; C069186:IE2 protein, Cytomegalovirus; C088646:IE3 protein, cytomegalovirus; D017874:Immediate-Early Proteins; D008562:Membrane Glycoproteins; D011498:Protein Precursors; D015534:Trans-Activators; C077406:UL115 protein, Human herpesvirus 5; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins; C071440:glycoprotein H, Cytomegalovirus; C499358:glycoprotein H, Human cytomegalovirus; C439068:glycoprotein O, cytomegalovirus