Human immunodeficiency virus type 1 protease microinjected into cultured human skin fibroblasts cleaves vimentin and affects cytoskeletal and nuclear architecture.
Human immunodeficiency virus type 1 protease microinjected into cultured human skin fibroblasts cleaves vimentin and affects cytoskeletal and nuclear architecture.
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发表时间：
1991-12
影响因子：
4
通讯作者：
B. Höner;R. Shoeman;P. Traub
中科院分区：
文献类型：
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作者：
B. Höner;R. Shoeman;P. Traub
在人皮肤成纤维细胞显微注射纯化的人类免疫缺陷病毒1型蛋白酶（HIV-1 PR），应力纤维丢失，核形态和核染色质凝聚的改变进行了观察。此后，波形蛋白中间丝（IF）网络崩溃。对微管无影响。虽然受影响的细胞从基质中丢失，但显示这些效应的细胞比例的最小估计值为50%。对单个细胞的观察表明，这些效应在很大程度上是不可逆的，并且是导致注射HIV-1 PR的细胞死亡的步骤。显微注射HIV-1 PR的各种稀释液后，观察到在对应力纤维和IF网络几乎没有或没有观察到影响的条件下，可检测到核形态和染色质凝聚的变化。用[35 S]甲硫氨酸标记并显微注射HIV-1 PR或BSA的细胞蛋白进行二维凝胶电泳。凝胶模式的主要差异是波形蛋白的量减少和新产品的出现与波形蛋白与HIV-1 PR体外处理后获得的裂解产物共迁移。因此，HIV-1 PR不仅能够在体内切割IF亚基蛋白，而且还可以催化其他细胞结构的改变。
In human skin fibroblasts microinjected with purified human immunodeficiency virus type 1 protease (HIV-1 PR), stress fibers were lost and alterations in nuclear morphology and condensation of nuclear chromatin were observed. Thereafter, the vimentin intermediate filament (IF) network collapsed. No effect was seen on the microtubules. While complicated by loss of affected cells from the substratum, a minimum estimate of the proportion of cells demonstrating these effects is 50%. Observation of single cells demonstrated that these effects were largely irreversible and were steps leading to the death of the HIV-1 PR-injected cells. After microinjection of various dilutions of the HIV-1 PR, it was observed that the changes in nuclear morphology and chromatin condensation were detectable under conditions where little or no effect was observed on both stress fibers and the IF network. Proteins of cells labelled with [35S]methionine and microinjected with either HIV-1 PR or BSA were subjected to two-dimensional gel electrophoresis. The major differences in the gel patterns were a diminution in the amount of vimentin and the appearance of novel products comigrating with cleavage products obtained after treatment of vimentin with HIV-1 PR in vitro. Thus, the HIV-1 PR is capable not only of cleaving IF subunit proteins in vivo, but also can catalyze alterations in other cellular structures.