EXPRESSION OF SPECIFIC MESSENGER-RNAS DURING ADIPOSE DIFFERENTIATION - IDENTIFICATION OF AN MESSENGER-RNA ENCODING A HOMOLOG OF MYELIN-P2 PROTEIN
EXPRESSION OF SPECIFIC MESSENGER-RNAS DURING ADIPOSE DIFFERENTIATION - IDENTIFICATION OF AN MESSENGER-RNA ENCODING A HOMOLOG OF MYELIN-P2 PROTEIN
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DOI：
10.1073/pnas.81.17.5468
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发表时间：
1984-01-01
期刊：
影响因子：
--
通讯作者：
KELLY, TJ
中科院分区：
文献类型：
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作者：
BERNLOHR, DA;ANGUS, CW;KELLY, TJ
为了鉴定和表征在小鼠3 T3-L1前脂肪细胞分化期间丰度增加的特定mRNA，从分离自分化的3 T3-L1脂肪细胞的poly（A）+ RNA构建c[互补]DNA文库。混合探针同位素比选择和RNA印迹分析已经确定了几个独特的cDNA克隆，代表mRNA的物种表达或独家或显着增加的水平在分化的细胞。对一个这样的克隆（pAL 422）的进一步表征揭示，仅在分化后才可检测到的相应mRNA具有大致相同的长度（600 ± 0.05）。150个碱基）作为cDNA插入片段（672个碱基）。pAL 422中cDNA插入片段的完整核苷酸序列显示了编码14.6 kDa [千道尔顿]的132个氨基酸多肽（422蛋白）的单个长开放阅读框。这些和其他结果表明，该cDNA可能代表mRNA的近全长拷贝。计算机辅助分析表明，422蛋白与兔和牛周围神经的髓鞘P2蛋白分别具有69%和64%的同源性，与大鼠肝脏和肠道的脂肪酸结合蛋白分别具有23%和30%的同源性。通过pAL 422 DNA选择的mRNA杂合体指导合适的基因的体外翻译。13 kDa多肽，并且该蛋白质通过抗牛髓磷脂P2的抗血清特异性免疫沉淀。显然，422蛋白是髓鞘P2的结构和可能的功能类似物。
To identify and characterize specific mRNA that increase in abundance during differentiation of mouse 3T3-L1 preadipocytes, a c[complementary]DNA library was constructed from poly(A)+ RNA isolated from differentiated 3T3-L1 adipocytes. Mixed probe isotope ratio selection and RNA blot analyses have identified several unique cDNA clones that represent mRNA species expressed either exclusively or at dramatically increased levels in differentiated cells. Further characterization of one such clone (pAL422) revealed that the corresponding mRNA, detectable only after differentiation, is approximately the same length (600 .+-. 150 bases) as the cDNA insert (672 bases). The complete nucleotide sequence of the cDNA insert in pAL422 revealed a single long open reading frame that encodes a 132 amino acid polypeptide (the 422 protein) of 14.6 kDa [kilodalton]. These and other results suggest that this cDNA may represent a nearly full-length copy of the mRNA. Computer-assisted analyses showed that the 422 protein shares 69% and 64% homology with myelin P2 proteins from rabbit and bovine peripheral nerves, respectively, as well as 23% and 30% homology with fatty-acid binding proteins from rat liver and intestine, respectively. The mRNA hybrid selected by pAL422 DNA directs the in vitro translation of an .apprxeq. 13 kDa polypeptide, and this protein is specifically immunoprecipitated by antiserum against bovine myelin P2. Evidently, the 422 protein is a structural, and possibly functional, analog of myelin P2.