Expression of Human Herpesvirus 6B Rep Within Infected Cells and Binding of Its Gene Product to the TATA-binding Protein in Vitro and in Vivo

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2000 Jun 14

PMID 10846093

Citations 16

Authors

Y Mori

P Dhepakson

T Shimamoto

K Ueda

Y Gomi

H Tani

Y Matsuura

K Yamanishi

Affiliations

Soon will be listed here.

Abstract

We have characterized the human herpesvirus 6B (HHV-6B) rep gene, which is a homologue of the adeno-associated virus type 2 rep and is unique in the herpesvirus family. Three transcripts, 9.0, 5.0, and 2. 7 kb (the major transcript), were detected by Northern blotting using an HHV-6B rep probe under late conditions. We investigated the expression kinetics of the rep gene using cycloheximide (CHX) and phosphonoformic acid (PFA), which are inhibitors of protein synthesis and viral DNA synthesis, respectively. The 5.2-kb transcript was mainly detected in the absence of protein biosynthesis upon infection, and none of the 9.0-, 5.0-, and 2.7-kb transcripts detected under the late conditions were detected in the presence of CHX and PFA. Sequences obtained from a cDNA library showed that the 5.0- and 2.7-kb transcripts were spliced from two and three exons, respectively, and the 2.7-kb transcript was more abundant. Immunohistochemistry using an antibody raised against the HHV-6 rep gene product (REP) revealed that REP was mainly present in the nucleus of MT-4 cells within 24 h after infection with HHV-6B. Using pull-down assays, coimmunoprecipitation, and a mammalian two hybrid system, we showed that HHV-6 REP binds to a transcription factor, human TATA-binding protein, through its N-terminal region.

Citing Articles

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Inhibition of DNA Repair Mechanisms and Induction of Apoptosis in Triple Negative Breast Cancer Cells Expressing the Human Herpesvirus 6 U94.

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Human Herpesvirus 6A and 6B inhibit in vitro angiogenesis by induction of Human Leukocyte Antigen G.

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Characterization of human herpesvirus 6A/B U94 as ATPase, helicase, exonuclease and DNA-binding proteins.

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References

Yamanishi K, Okuno T, Shiraki K, Takahashi M, Kondo T, Asano Y . Identification of human herpesvirus-6 as a causal agent for exanthem subitum. Lancet. 1988; 1(8594):1065-7. DOI: 10.1016/s0140-6736(88)91893-4. View

Braun D, Dominguez G, Pellett P . Human herpesvirus 6. Clin Microbiol Rev. 1997; 10(3):521-67. PMC: 172933. DOI: 10.1128/CMR.10.3.521. View

Thomson B, Weindler F, Gray D, Schwaab V, Heilbronn R . Human herpesvirus 6 (HHV-6) is a helper virus for adeno-associated virus type 2 (AAV-2) and the AAV-2 rep gene homologue in HHV-6 can mediate AAV-2 DNA replication and regulate gene expression. Virology. 1994; 204(1):304-11. DOI: 10.1006/viro.1994.1535. View

Hahn S . The role of TAFs in RNA polymerase II transcription. Cell. 1998; 95(5):579-82. DOI: 10.1016/s0092-8674(00)81625-6. View

Weger S, Wendland M, Kleinschmidt J, Heilbronn R . The adeno-associated virus type 2 regulatory proteins rep78 and rep68 interact with the transcriptional coactivator PC4. J Virol. 1998; 73(1):260-9. PMC: 103830. DOI: 10.1128/JVI.73.1.260-269.1999. View

Lee T, Young R . Regulation of gene expression by TBP-associated proteins. Genes Dev. 1998; 12(10):1398-408. DOI: 10.1101/gad.12.10.1398. View

Kyostio S, Wonderling R, Owens R . Negative regulation of the adeno-associated virus (AAV) P5 promoter involves both the P5 rep binding site and the consensus ATP-binding motif of the AAV Rep68 protein. J Virol. 1995; 69(11):6787-96. PMC: 189590. DOI: 10.1128/JVI.69.11.6787-6796.1995. View

Antoni B, Rabson A, MILLER I, Trempe J, Chejanovsky N, Carter B . Adeno-associated virus Rep protein inhibits human immunodeficiency virus type 1 production in human cells. J Virol. 1991; 65(1):396-404. PMC: 240530. DOI: 10.1128/JVI.65.1.396-404.1991. View

Carrigan D, Knox K, Tapper M . Suppression of human immunodeficiency virus type 1 replication by human herpesvirus-6. J Infect Dis. 1990; 162(4):844-51. DOI: 10.1093/infdis/162.4.844. View

10.

Mori Y, Yagi H, Shimamoto T, Isegawa Y, Sunagawa T, Inagi R . Analysis of human herpesvirus 6 U3 gene, which is a positional homolog of human cytomegalovirus UL 24 gene. Virology. 1998; 249(1):129-39. DOI: 10.1006/viro.1998.9305. View

11.

TORELLI G, Barozzi P, Marasca R, Cocconcelli P, Merelli E, Ceccherini-Nelli L . Targeted integration of human herpesvirus 6 in the p arm of chromosome 17 of human peripheral blood mononuclear cells in vivo. J Med Virol. 1995; 46(3):178-88. DOI: 10.1002/jmv.1890460303. View

12.

Lopez C, Pellett P, Stewart J, Goldsmith C, Sanderlin K, Black J . Characteristics of human herpesvirus-6. J Infect Dis. 1988; 157(6):1271-3. DOI: 10.1093/infdis/157.6.1271. View

13.

Hoffmann A, Roeder R . Purification of his-tagged proteins in non-denaturing conditions suggests a convenient method for protein interaction studies. Nucleic Acids Res. 1991; 19(22):6337-8. PMC: 329157. DOI: 10.1093/nar/19.22.6337. View

14.

Labow M, Hermonat P, Berns K . Positive and negative autoregulation of the adeno-associated virus type 2 genome. J Virol. 1986; 60(1):251-8. PMC: 253923. DOI: 10.1128/JVI.60.1.251-258.1986. View

15.

Martin D, Subler M, Munoz R, Brown D, Deb S, Deb S . p53 and SV40 T antigen bind to the same region overlapping the conserved domain of the TATA-binding protein. Biochem Biophys Res Commun. 1993; 195(1):428-34. DOI: 10.1006/bbrc.1993.2061. View

16.

Matsuura Y, Possee R, Overton H, Bishop D . Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. J Gen Virol. 1987; 68 ( Pt 5):1233-50. DOI: 10.1099/0022-1317-68-5-1233. View

17.

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

18.

Hermonat P . Down-regulation of the human c-fos and c-myc proto-oncogene promoters by adeno-associated virus Rep78. Cancer Lett. 1994; 81(2):129-36. DOI: 10.1016/0304-3835(94)90193-7. View

19.

Gompels U, Nicholas J, Lawrence G, Jones M, Thomson B, Martin M . The DNA sequence of human herpesvirus-6: structure, coding content, and genome evolution. Virology. 1995; 209(1):29-51. DOI: 10.1006/viro.1995.1228. View

20.

Heilbronn R, Burkle A, Stephan S, zur Hausen H . The adeno-associated virus rep gene suppresses herpes simplex virus-induced DNA amplification. J Virol. 1990; 64(6):3012-8. PMC: 249485. DOI: 10.1128/JVI.64.6.3012-3018.1990. View