Three sites of the hepatitis B virus X protein cooperatively interact with cellular proteins.
The X protein of hepatitis B virus is known to be a trans-activator of viral and cellular genes and to be a serine protease inhibitor as well. X protein has no DNA-binding activity, but is postulated to exert its trans-activation function by interacting with cellular proteins. To investigate interaction sites of X protein with cellular proteins, we carried out an immunoprecipitation inhibition assay using several different anti-X antibodies in the presence or absence of cellular proteins. Results elucidated three separate sites (aa 65-72, aa 105-115, and aa 131-142; U22, X1, and Z44 sites, respectively) of the X protein that cooperatively interacted with cellular proteins. Analyses with a series of mutant X proteins also supported the interactions at the U22, X1, and Z44 sites. Based on the CAT activity assay, the essential regions for the trans-activation function of X protein overlapped with these three interaction sites. Furthermore, these interaction sites also coincide with the structures necessary for the serine protease inhibitor activity. Thus, the trans-activation function and serine protease inhibitor activity of X protein may be exerted by interaction with cellular proteins through at least these three sites.
Department of Gene Research, Cancer Institute (JFCR), Tokyo, Japan.
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Takada
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Koike
D000494:Allosteric Regulation; D000495:Allosteric Site; D000595:Amino Acid Sequence; D000818:Animals; D006515:Hepatitis B virus / Q000737:chemistry*; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D011485:Protein Binding; D014176:Protein Biosynthesis; D011817:Rabbits; D012697:Serine Endopeptidases / Q000378:metabolism; D015534:Trans-Activators / Q000737:chemistry* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D054334:Viral Regulatory and Accessory Proteins
D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; D012697:Serine Endopeptidases