首页> 美国卫生研究院文献>Journal of Virology >The p12I p13II and p30II proteins encoded by human T-cell leukemia/lymphotropic virus type I open reading frames I and II are localized in three different cellular compartments.
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The p12I p13II and p30II proteins encoded by human T-cell leukemia/lymphotropic virus type I open reading frames I and II are localized in three different cellular compartments.

机译:由人类T细胞白血病/淋巴病毒I型开放阅读框I和II编码的p12Ip13II和p30II蛋白位于三个不同的细胞区室中。

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摘要

Three protein isoforms are encoded by the human T-cell leukemia/lymphotropic virus type I pX region open reading frames (ORF) I and II through alternative splicing. Both the singly and doubly spliced mRNAs from ORF I encode a single 12-kDa protein (p12I), whereas two distinct proteins of 13 kDa (p13II) and 30 kDa (p30II) are encoded from the ORF II alternatively spliced mRNA. Because the p12I protein is very hydrophobic and poorly immunogenic, we genetically engineered its cDNA by adding a short stretch of amino acids from the highly immunogenic epitope HA1 of influenza virus or the AU1 epitope of bovine papillomavirus. The HA1 epitope was also added to the p13II and p30II proteins, albeit rabbit immune sera raised against synthetic peptides were also available. To determine in which cellular compartments these proteins reside, we transfected the tagged and wild-type cDNAs in HeLa/Tat cells and studied their localization by indirect immunofluorescence. The p12I protein was identified in the cellular endomembranes and, particularly, in the perinuclear area. p13II and p30II were found in the nuclei and nucleoli of the transfected cells, respectively. The presence of the HA1 epitope at the carboxy terminus of p13II and p30II did not interfere with their cellular localization, since the rabbit immune sera demonstrated their presence in the same cellular compartments when the untagged proteins were expressed. The defined localization of these proteins in specific cellular compartments warrants further study of their function.
机译:人类T细胞白血病/淋巴病毒I型pX区开放阅读框(ORF)I和II通过交替剪接编码三种蛋白质同工型。 ORF I的单剪接和双剪接的mRNA均编码单个12-kDa蛋白(p12I),而ORF II剪接的mRNA则编码了13 kDa(p13II)和30 kDa(p30II)的两种截然不同的蛋白。由于p12I蛋白疏水性强且免疫原性差,因此我们通过从流感病毒的高度免疫原性表位HA1或牛乳头瘤病毒AU1表位中添加一小段氨基酸对基因进行了遗传工程改造。 HA1表位也添加到了p13II和p30II蛋白中,尽管也有针对合成肽的兔免疫血清。为了确定这些蛋白质驻留在哪些细胞区室中,我们在HeLa / Tat细胞中转染了标记的和野生型cDNA,并通过间接免疫荧光研究了它们的定位。在细胞内膜中,尤其是在核周区域中鉴定出p12I蛋白。在转染细胞的细胞核和核仁中分别发现了p13II和p30II。在p13II和p30II的羧基末端存在HA1表位不会干扰它们的细胞定位,因为当表达未标记的蛋白时,兔的免疫血清证明了它们在相同的细胞区室中的存在。这些蛋白质在特定细胞区室中的确定定位,有待进一步研究其功能。

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