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首页> 美国卫生研究院文献>Journal of Virology >Impaired Maturation of Mouse Mammary Tumor Virus Precursor Polypeptides in Lymphoid Leukemia Cells Producing Intracytoplasmic A Particles and No Extracellular B-Type Virions
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Impaired Maturation of Mouse Mammary Tumor Virus Precursor Polypeptides in Lymphoid Leukemia Cells Producing Intracytoplasmic A Particles and No Extracellular B-Type Virions

机译：淋巴样白血病细胞中小鼠乳腺肿瘤病毒前体多肽的成熟受损产生胞浆内A粒子没有细胞外B型病毒体。

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Processing of polypeptides of the mouse mammary tumor virus, a type B retrovirus, was investigated in a transplanted thymic lymphoma cell line of the GR strain (GRSL). This cell line was maintained in vivo in ascites form and in vitro as a suspension culture. GRSL cells produce clusters of intracytoplasmic A particles and are virtually deficient in the production of mature extracellular B-type particles. As control, a mammary tumor cell line of the same mouse strain capable of complete virion synthesis was used. The kinetics of viral polypeptide synthesis were studied by pulse labeling with various isotopes (including 35S and 32P), followed by immunoprecipitation of cell lysates with monospecific antisera to the major mouse mammary tumor virus gag and env proteins, p27 and gp52, respectively. Both the primary gag and env precursor polypeptides were synthesized in the GRSL cells, but their conversion into viral proteins was impaired. The major gag precursor, Pr73gag, was stable over a period of 8 h, and mature viral core polypeptides could not be detected. Also, the highly phosphorylated intermediates in the proteolytic processing of Pr73gag in virus-producing cells were absent in GRSL cells. By immunoprecipitation, Pr73gag was detected in a GRSL particle fraction with the density of intracytoplasmic A particles. The precursor for envelope proteins, Pr73env, was turned over without the generation of mature viral envelope components gp52 and gp36. The in vivo-transplanted ascites GRSL cells, however, were shown to express gp52 on the cell surface together with a 73,000-dalton polypeptide, as indicated by cell surface iodination and immunoprecipitation.
机译：在移植的GR株胸腺淋巴瘤细胞系（GRSL）中研究了小鼠乳腺肿瘤病毒B型逆转录病毒多肽的加工过程。该细胞系以腹水形式体内维持，并在体外以悬浮培养物形式维持。 GRSL细胞产生胞浆内A颗粒簇，实际上不足以产生成熟的细胞外B型颗粒。作为对照，使用能够完成病毒体合成的相同小鼠品系的乳腺肿瘤细胞系。通过用各种同位素（包括 35 S和 32 P）进行脉冲标记研究病毒多肽合成的动力学，然后用单特异性抗血清对主要小鼠免疫沉淀细胞裂解物乳腺肿瘤病毒gag和env蛋白分别为p27和gp52。主要的gag和env前体多肽都在GRSL细胞中合成，但是它们转化为病毒蛋白的过程受到损害。主要gag前体Pr73 gag 在8小时内稳定，并且无法检测到成熟的病毒核心多肽。同样，在GRSL细胞中不存在病毒产生细胞中Pr73 gag 的蛋白水解过程中高度磷酸化的中间体。通过免疫沉淀，在具有胞浆内A颗粒密度的GRSL颗粒级分中检测到Pr73 gag 。包膜蛋白的前体Pr73 env 被移交而没有生成成熟的病毒包膜成分gp52和gp36。然而，如细胞表面碘化和免疫沉淀所表明，体内移植的腹水GRSL细胞与73,000道尔顿多肽一起在细胞表面表达gp52。

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