Kalcheva N, Shafit-Zagardo B
Department of Pathology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.
J Neurochem. 1995 Oct;65(4):1472-80. doi: 10.1046/j.1471-4159.1995.65041472.x.
Three unique 5' untranslated regions (UTRs) have been characterized for human microtubule-associated protein-2 (MAP-2) transcripts. All three UTRs shared a common 171-bp sequence adjacent to the MAP-2 coding region and then diverged upstream. The size of the unique upstream sequence was 281, 146, or 104 bp. PCR of genomic DNA demonstrated that the 5' UTRs span multiple exons. The unique region of the UTRs recognizes a 9.5- and a 6-kb MAP-2 transcript in poly(A)+ mRNA isolated from human MSN cells, and PCR analysis demonstrated that each unique UTR is contained in multiple high- and low-molecular-weight MAP-2 transcripts. Reverse transcription-PCR (RT-PCR) performed on MSN mRNA isolated from polysomes demonstrated that all three of the UTRs contained within multiple MAP-2 transcripts were associated with polysomes and hence translated. RT-PCR from human fetal spinal cord and adult brain mRNA demonstrated that all of the UTRs are expressed at these developmental time points.
已对人类微管相关蛋白2(MAP-2)转录本的三个独特的5'非翻译区(UTR)进行了表征。所有这三个UTR在与MAP-2编码区相邻处共享一个171bp的共同序列,然后在其上游出现分歧。独特的上游序列大小分别为281bp、146bp或104bp。基因组DNA的PCR结果表明,5'UTR跨越多个外显子。UTR的独特区域在从人MSN细胞分离的多聚腺苷酸加尾(poly(A)+)mRNA中识别出9.5kb和6kb的MAP-2转录本,并且PCR分析表明每个独特的UTR都包含在多个高分子量和低分子量的MAP-2转录本中。对从多核糖体分离的MSN mRNA进行的逆转录PCR(RT-PCR)表明,多个MAP-2转录本中包含的所有三个UTR都与多核糖体相关,因此是可翻译的。来自人类胎儿脊髓和成人脑mRNA的RT-PCR表明,所有UTR在这些发育时间点均有表达。