Characterization of equine herpesvirus type 1 immediate early proteins.
Characterization of equine herpesvirus type 1 immediate early proteins.
复制标题
马疱疹病毒 1 型立即早期蛋白的表征。
DOI：
10.1016/0042-6822(88)90297-8
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发表时间：
1988
期刊：
影响因子：
3.7
通讯作者：
O'Callaghan,DJ
中科院分区：
文献类型：
--
作者：
Caughman,GB;Robertson,AT;Gray,WL;Sullivan,DC;O'Callaghan,DJ
EHV-1即刻早期(IE)基因在溶血性感染中的表达导致4种高摩尔质量的即刻早期多肽(IEP)的产生，分别为IE1、IE2、IE2和IE4；然而，IE的转录仅限于合成单个6-kb的mRNA。总而言之，这些发现提出了一个问题，即这四个IEP是否是同一基因的相关产物。在本研究中，从结构相似性、抗原相关性和合成后修饰三个方面对免疫球蛋白进行了表征。IE1是最丰富的IEP，在IE条件下用[35S]蛋氨酸或14C标记的氨基酸混合物标记的感染兔肾细胞中，IE1约占IEP掺入放射性标记的80%。IE1也是主要的磷酸化物种。用葡萄球菌V8酶对分离的放射性标记的IEP条带进行有限的蛋白水解酶消化，在SDS-PAGE中得到几乎相同的片段图谱，用糜蛋白酶和N-氯丁二酰亚胺进行消化也是如此。在免疫印迹试验中，针对4个分离的IEP分别制备的单特异性兔抗血清与所有IEP物种发生反应。脉冲追逐实验表明，在15分钟的脉冲后立即可以检测到所有的IEP，并且在随后的追逐期间IEP轮廓发生了几次变化。因此，EHV-1免疫球蛋白在结构和抗原性上密切相关，似乎要么同时产生，要么被加工成立即产生单个形式。
EHV-1 immediate early (IE) gene expression in lytic infection results in the production of four high mol wt immediate early polypeptides (IEPs), designated IE1, IE2, IE2, and IE4; however, IE transcription is limited to the synthesis of a single 6-kb mRNA. Together, these findings raised questions as to whether the four IEPs were related products of the same gene. In the present study the IEPs were characterized with respect to their structural similarities, antigenic relatedness, and postsynthetic modifications. IE1 was the most abundant IEP, in that it accounted for approximately 80% of the IEP-incorporated radiolabel in infected rabbit kidney cells labeled under IE conditions with [35S]methionine or14C-labeled amino acid mixtures. IE1 also was the major phosphorylated species. Limited proteolytic digestion of isolated radiolabeled IEP bands with Staph V8 protease yielded virtually identical fragment profiles in SDS-PAGE, as did digestions with chymotrypsin andN-chlorosuccinimide. Monospecific rabbit antisera raised against each of the four isolated IEPs reacted with all the IEP species in immunoblotting assays. Pulse-chase experiments indicated that all the IEPs were detectable immediately after a 15-min pulse and that several alterations in the IEP profile occurred during subsequent chase periods. Thus, the EHV-1 IEPs are closely related structurally and antigenically and appeared to be either produced simultaneously or processed to yield the individual forms immediately.