Functional analysis of a carboxyl-terminal phosphorylation mutant of the bovine papillomavirus E1 protein.
Functional analysis of a carboxyl-terminal phosphorylation mutant of the bovine papillomavirus E1 protein.
复制标题
牛乳头瘤病毒 E1 蛋白羧基末端磷酸化突变体的功能分析。
DOI：
10.1006/jmbi.2001.5375
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Hella,Maria
中科院分区：
文献类型：
--
作者：
Lentz,Michael;Zanardi,Thomas;Filzen,Robyn;Carter,Jena;Hella,Maria
乳头瘤病毒 E1 蛋白对于病毒 DNA 复制至关重要，E1 的磷酸化似乎可以调节蛋白功能和 DNA 复制。牛乳头瘤病毒E1的丝氨酸584位于类似于CK2共有位点的保守基序中，并且在体外被CK2磷酸化。在瞬时复制测定中，丝氨酸 584 突变为丙氨酸会消除病毒基因组的复制。野生型和突变型 E1 蛋白由重组杆状病毒表达，用于评估氨基酸 584 取代的生化功能。解旋酶活性、E1 与病毒 E2 蛋白的结合以及与细胞 DNA 聚合酶 α-引物酶的结合在突变蛋白中均不受影响。在突变体中，E1 与病毒复制起点 DNA 序列的结合减少，但并未消除。该蛋白的羧基末端区域似乎在调节 E1 功能中发挥作用，并为乳头瘤病毒 DNA 复制控制的复杂图景增添了色彩。
The papillomavirus E1 protein is essential for viral DNA replication, and phosphorylation of E1 appears to regulate protein function and DNA replication. Serine 584 of bovine papillomavirus E1 is in a conserved motif resembling a CK2 consensus site, and is phosphorylated by CK2 in vitro. Mutation of serine 584 to alanine eliminates replication of the viral genome in transient replication assays. Wild-type and mutant E1 proteins were expressed from recombinant baculoviruses and used to assess biochemical functions of the amino acid 584 substitution. Helicase enzyme activity, E1 binding to the viral E2 protein and to cellular DNA polymerase alpha-primase were all unaffected in the mutant protein. Binding of E1 to viral replication origin DNA sequences was reduced in the mutant, but not eliminated. The carboxyl-terminal region of the protein appears to play a role in regulating E1 function, and adds to a complex picture emerging for papillomavirus DNA replication control.