Identification and characterization of glycoprotein gh of human herpesvirus-6

Gao Qian, Charles Wood, Bala Chandran

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Sequence analysis of an 8-kb BamHI-G DNA fragment from human herpesvirus-6 variant A strain GS (HHV-6A(GS)) has identified a gene encoding a glycoprotein homologous to glycoprotein gH of other herpesviruses and this gene was designated as HHV-6 gH (Josephs et al., J. Virol. 65, 5597). The open reading frame (ORF) of HHV-6A(GS) gH gene was amplified by polymerase chain reaction and was expressed as a trpE fusion protein in bacteria. Stable fusion protein was made only with constructs containing amino acids 15 to 469 of the ORF and this fusion protein was used to raise rabbit antisera. In the in vitro transcription-translation analysis of the gH gene, addition of microsomal membranes resulted in the processing of an unprocessed precursor to a polypeptide product of about 108 K. These in vitro-synthesized precursor and product forms were immunoprecipitated by the rabbit antibodies against the gH fusion protein. Rabbit anti-gH antibodies neutralized the infectivity of both HHV-6 variant A strain GS and variant B strain Z-29, and immunoprecipitation reactions identified a virion envelope-associated 102 K polypeptide as the glycoprotein gH of HHV-6 variants A and B. In addition to the 102 K glycoprotein, nonglycosylated polypeptides of about 58 and 164 K with different partial peptide maps were also consistently coimmunoprecipitated from (35S]methionine-labeled HHV-6A(GS)-infected cells, but not from HHV-6B(Z-29)-infected cells. The similarity in the molecular weights of glycoprotein gH among the two strains belonging to the two variant groups of HHV-6 and neutralization of infectivity of both groups by rabbit antibodies against HHV-6A(GS) gH suggests conserved functions of glycoprotein gH among the two variant groups of HHV-6.

Original languageEnglish (US)
Pages (from-to)380-386
Number of pages7
JournalVirology
Volume194
Issue number1
DOIs
StatePublished - May 1993

Fingerprint

Human Herpesvirus 6
Glycoproteins
Rabbits
Peptides
Open Reading Frames
Genes
Proteins
herpesvirus 6 glycoprotein H
Antibodies
Herpesviridae
Immunoprecipitation
Methionine
Virion
Sequence Analysis
Immune Sera
Anti-Idiotypic Antibodies
Molecular Weight
Bacteria
Amino Acids
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Virology

Cite this

Identification and characterization of glycoprotein gh of human herpesvirus-6. / Qian, Gao; Wood, Charles; Chandran, Bala.

In: Virology, Vol. 194, No. 1, 05.1993, p. 380-386.

Research output: Contribution to journalArticle

Qian, Gao ; Wood, Charles ; Chandran, Bala. / Identification and characterization of glycoprotein gh of human herpesvirus-6. In: Virology. 1993 ; Vol. 194, No. 1. pp. 380-386.
@article{714cb4a47e164b5da1b84c1177a0ceb6,
title = "Identification and characterization of glycoprotein gh of human herpesvirus-6",
abstract = "Sequence analysis of an 8-kb BamHI-G DNA fragment from human herpesvirus-6 variant A strain GS (HHV-6A(GS)) has identified a gene encoding a glycoprotein homologous to glycoprotein gH of other herpesviruses and this gene was designated as HHV-6 gH (Josephs et al., J. Virol. 65, 5597). The open reading frame (ORF) of HHV-6A(GS) gH gene was amplified by polymerase chain reaction and was expressed as a trpE fusion protein in bacteria. Stable fusion protein was made only with constructs containing amino acids 15 to 469 of the ORF and this fusion protein was used to raise rabbit antisera. In the in vitro transcription-translation analysis of the gH gene, addition of microsomal membranes resulted in the processing of an unprocessed precursor to a polypeptide product of about 108 K. These in vitro-synthesized precursor and product forms were immunoprecipitated by the rabbit antibodies against the gH fusion protein. Rabbit anti-gH antibodies neutralized the infectivity of both HHV-6 variant A strain GS and variant B strain Z-29, and immunoprecipitation reactions identified a virion envelope-associated 102 K polypeptide as the glycoprotein gH of HHV-6 variants A and B. In addition to the 102 K glycoprotein, nonglycosylated polypeptides of about 58 and 164 K with different partial peptide maps were also consistently coimmunoprecipitated from (35S]methionine-labeled HHV-6A(GS)-infected cells, but not from HHV-6B(Z-29)-infected cells. The similarity in the molecular weights of glycoprotein gH among the two strains belonging to the two variant groups of HHV-6 and neutralization of infectivity of both groups by rabbit antibodies against HHV-6A(GS) gH suggests conserved functions of glycoprotein gH among the two variant groups of HHV-6.",
author = "Gao Qian and Charles Wood and Bala Chandran",
year = "1993",
month = "5",
doi = "10.1006/viro.1993.1272",
language = "English (US)",
volume = "194",
pages = "380--386",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Identification and characterization of glycoprotein gh of human herpesvirus-6

AU - Qian, Gao

AU - Wood, Charles

AU - Chandran, Bala

PY - 1993/5

Y1 - 1993/5

N2 - Sequence analysis of an 8-kb BamHI-G DNA fragment from human herpesvirus-6 variant A strain GS (HHV-6A(GS)) has identified a gene encoding a glycoprotein homologous to glycoprotein gH of other herpesviruses and this gene was designated as HHV-6 gH (Josephs et al., J. Virol. 65, 5597). The open reading frame (ORF) of HHV-6A(GS) gH gene was amplified by polymerase chain reaction and was expressed as a trpE fusion protein in bacteria. Stable fusion protein was made only with constructs containing amino acids 15 to 469 of the ORF and this fusion protein was used to raise rabbit antisera. In the in vitro transcription-translation analysis of the gH gene, addition of microsomal membranes resulted in the processing of an unprocessed precursor to a polypeptide product of about 108 K. These in vitro-synthesized precursor and product forms were immunoprecipitated by the rabbit antibodies against the gH fusion protein. Rabbit anti-gH antibodies neutralized the infectivity of both HHV-6 variant A strain GS and variant B strain Z-29, and immunoprecipitation reactions identified a virion envelope-associated 102 K polypeptide as the glycoprotein gH of HHV-6 variants A and B. In addition to the 102 K glycoprotein, nonglycosylated polypeptides of about 58 and 164 K with different partial peptide maps were also consistently coimmunoprecipitated from (35S]methionine-labeled HHV-6A(GS)-infected cells, but not from HHV-6B(Z-29)-infected cells. The similarity in the molecular weights of glycoprotein gH among the two strains belonging to the two variant groups of HHV-6 and neutralization of infectivity of both groups by rabbit antibodies against HHV-6A(GS) gH suggests conserved functions of glycoprotein gH among the two variant groups of HHV-6.

AB - Sequence analysis of an 8-kb BamHI-G DNA fragment from human herpesvirus-6 variant A strain GS (HHV-6A(GS)) has identified a gene encoding a glycoprotein homologous to glycoprotein gH of other herpesviruses and this gene was designated as HHV-6 gH (Josephs et al., J. Virol. 65, 5597). The open reading frame (ORF) of HHV-6A(GS) gH gene was amplified by polymerase chain reaction and was expressed as a trpE fusion protein in bacteria. Stable fusion protein was made only with constructs containing amino acids 15 to 469 of the ORF and this fusion protein was used to raise rabbit antisera. In the in vitro transcription-translation analysis of the gH gene, addition of microsomal membranes resulted in the processing of an unprocessed precursor to a polypeptide product of about 108 K. These in vitro-synthesized precursor and product forms were immunoprecipitated by the rabbit antibodies against the gH fusion protein. Rabbit anti-gH antibodies neutralized the infectivity of both HHV-6 variant A strain GS and variant B strain Z-29, and immunoprecipitation reactions identified a virion envelope-associated 102 K polypeptide as the glycoprotein gH of HHV-6 variants A and B. In addition to the 102 K glycoprotein, nonglycosylated polypeptides of about 58 and 164 K with different partial peptide maps were also consistently coimmunoprecipitated from (35S]methionine-labeled HHV-6A(GS)-infected cells, but not from HHV-6B(Z-29)-infected cells. The similarity in the molecular weights of glycoprotein gH among the two strains belonging to the two variant groups of HHV-6 and neutralization of infectivity of both groups by rabbit antibodies against HHV-6A(GS) gH suggests conserved functions of glycoprotein gH among the two variant groups of HHV-6.

UR - http://www.scopus.com/inward/record.url?scp=0027220977&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027220977&partnerID=8YFLogxK

U2 - 10.1006/viro.1993.1272

DO - 10.1006/viro.1993.1272

M3 - Article

C2 - 8386882

AN - SCOPUS:0027220977

VL - 194

SP - 380

EP - 386

JO - Virology

JF - Virology

SN - 0042-6822

IS - 1

ER -