Identification of glycoprotein gpTRL10 as a structural component of human cytomegalovirus.
Human cytomegalovirus (HCMV) has a coding capacity for glycoproteins which far exceeds that of other herpesviruses. Few of these proteins have been characterized. We have investigated the gene product(s) of reading frame 10, which is present in both the internal and terminal repeat regions of HCMV strain AD169 and only once in clinical isolates. The putative protein product is a 171-amino-acid glycoprotein with a theoretical mass of 20.5 kDa. We characterized the protein encoded by this reading frame in the laboratory strain AD169 and a recent isolate, TB40E. The results from both strains were comparable. Northern blot analyses showed that the gene was transcribed with early/late kinetics. Two proteins of 22 and 23.5-kDa were detected in virus-infected cells and in cells transiently expressing recombinant TRL10. Both forms contained only high-mannose-linked carbohydrate modifications. In addition, virus-infected cells expressed small amounts of the protein modified with complex N-linked sugars. Image analysis localized transiently expressed TRL10 to the endoplasmic reticulum. Immunoblot analyses as well as immunoelectron microscopy of purified virions demonstrated that TRL10 represents a structural component of the virus particle. Immunoblot analysis in the absence of reducing agents indicated that TRL10, like the other HCMV envelope glycoproteins, is present in a disulfide-linked complex. Sequence analysis of the TRL10 coding region in nine low-passage clinical isolates revealed strain-specific variation. In summary, the protein product of the TRL10 open reading frame represents a novel structural glycoprotein of HCMV and was termed gpTRL10.
Spaderna
S
S
Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, 91054 Erlangen, Germany.
Blessing
H
H
Bogner
E
E
Britt
W
W
Mach
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007424:Intracellular Fluid / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020079:Terminal Repeat Sequences; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
D004279:DNA, Viral; D006023:Glycoproteins; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; C446359:TRL10 protein, Human cytomegalovirus; D014759:Viral Envelope Proteins