Hou X, Foley S, Cueto M, Robinson M A
Laboratory of Immunogenetics, Twinbrook II Facility, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 12441 Parklawn Drive, Rockville, Maryland, 20852, USA.
Virology. 2000 Nov 10;277(1):127-35. doi: 10.1006/viro.2000.0604.
Interactions between the Human T-cell leukemia virus type I (HTLV-I) gene product p13(II) and cellular proteins were investigated using the yeast two-hybrid system. Variant forms of p13(II) were derived from two HTLV-I molecular clones, K30p and K34p, that differ in both virus production and in vivo and in vitro infectivity. Two nucleotide differences between the p13 from K30p (p13K30) and K34p (p13K34) result in a Trp-Arg substitution at amino acid 17 and the truncation of the 25 carboxyl-terminal residues of p13K34. A cDNA library from an HTLV-I-infected rabbit T-cell line was screened with p13K30 and p13K34 as bait. Products of two cDNA clones, C44 and C254, interacted with p13K34 but not with p13K30. Interactions were further confirmed using the GST-fusion protein coprecipitation assay. Sequence analysis of C44 and C254 cDNA clones revealed similarities to members of the nucleoside monophosphate kinase superfamily and actin-binding protein 280, respectively. Further analysis of the function of these two proteins and the consequence of their interaction with p13 may help elucidate a role for p13 in virus production, infectivity, or the pathogenesis of HTLV-I.
利用酵母双杂交系统研究了人类I型T细胞白血病病毒（HTLV-I）基因产物p13(II)与细胞蛋白之间的相互作用。p13(II)的变异形式源自两个HTLV-I分子克隆K30p和K34p，这两个克隆在病毒产生以及体内和体外感染性方面均存在差异。K30p的p13（p13K30）与K34p的p13（p13K34）之间的两个核苷酸差异导致第17位氨基酸处色氨酸-精氨酸替换，以及p13K34的25个羧基末端残基截短。以p13K30和p13K34作为诱饵，筛选来自HTLV-I感染的兔T细胞系的cDNA文库。两个cDNA克隆C44和C254的产物与p13K34相互作用，但不与p13K30相互作用。使用GST融合蛋白共沉淀试验进一步证实了相互作用。对C44和C254 cDNA克隆的序列分析分别显示与核苷单磷酸激酶超家族成员和肌动蛋白结合蛋白280相似。对这两种蛋白质的功能及其与p13相互作用的后果进行进一步分析，可能有助于阐明p13在病毒产生、感染性或HTLV-I发病机制中的作用。
