Gallina A, Mantoan G, Rindi G, Milanesi G
Istituto di Genetica biochimica ed Evoluzionistica, CNR, Pavia, Italy.
Biochem Biophys Res Commun. 1994 Nov 15;204(3):1031-8. doi: 10.1006/bbrc.1994.2566.
HIV-1 Gag protein intracellular transport and budding was investigated by altering the sequence of the MA domain, which directly bears an essential N-terminal myristyl adduct and forms the viral matrix after Gag proteolysis in mature virions. We found that removal of a substantial MA internal segment did not abolish the assembly and budding of Gag particles, but rather diverted these events to intracellular cisternae. The internally deleted Gag was further modified by substituting either of two heterologous myristylated N-termini for the natural one: amino acids 1-12 from v-Src oncoprotein (for which a membrane-bound intracellular receptor has been postulated), or amino acids 1-12 from Poliovirus polyprotein (for which no membrane-targeting function has been demonstrated). Both Src-Gag and Polio-Gag chimerae exhibited transport and processing characteristics similar to those of the MA-deleted Gag. These results are discussed with respect to the possible transport pathway of HIV-1 Gag.
通过改变基质(MA)结构域的序列来研究HIV-1 Gag蛋白的细胞内运输和出芽,MA结构域直接带有一个必需的N端肉豆蔻酰加合物,并在成熟病毒粒子中Gag蛋白水解后形成病毒基质。我们发现,去除大量的MA内部片段并没有消除Gag颗粒的组装和出芽,而是将这些事件转移到细胞内的池状结构中。通过用两个异源肉豆蔻酰化的N端之一替换天然的N端,对内部缺失的Gag进行了进一步修饰:来自v-Src癌蛋白的1-12位氨基酸(假定其具有膜结合的细胞内受体),或来自脊髓灰质炎病毒多聚蛋白的1-12位氨基酸(尚未证明其具有膜靶向功能)。Src-Gag和脊髓灰质炎-Gag嵌合体均表现出与缺失MA的Gag相似的运输和加工特性。针对HIV-1 Gag可能的运输途径对这些结果进行了讨论。