PRAC: A novel small nuclear protein that is specifically expressed in human prostate and colon

PRAC: A novel small nuclear protein that is specifically expressed in human prostate and colon
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DOI:
10.1002/pros.1055
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发表时间:
2001-05-01
期刊:
影响因子:
2.8
通讯作者:
Pastan, I
Pastan, I
中科院分区:
医学3区
文献类型:
--
作者:
Liu, XF;Olsson, P;Pastan, I

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背景人类表达序列标签(dbEST)数据库为组织特异性基因的鉴定提供了潜在的来源。该数据库包含源自特定肿瘤、器官或细胞类型的cDNA文库的序列。在本报告中,我们使用EST数据库来鉴定PRAC,这是一种在人类前列腺、前列腺癌、直肠和远端结肠中特异性表达的新基因。方法。使用基于计算机的分析,序列同源EST的集群被确定为包含仅来自人前列腺cDNA文库的EST。采用多组织RNA斑点杂交和RT-PCR检测组织特异性。利用北方印迹分析、RACE-PCR、引物延伸和western blot鉴定PRAC转录物和蛋白Mas。PRAC编码一种382个核苷酸的RNA,存在于前列腺、直肠、远端结肠和三种前列腺癌细胞系中; LNCaP、PC-3和DU 145。该转录本编码6 kDa的核蛋白。PRAC基因定位于17号染色体17 q21,Hoxb-13基因下游约4kbp处。我们的数据证明EST数据库可以成为发现前列腺特异性基因的有用工具。核定位,鉴定潜在的磷酸化位点,并可能与Hoxb-13基因的共转录表明,PRAC可能在细胞核中的调节作用。前列腺47:125-131,2001年。(C)2001 Wiley-Liss,Inc.
BACKGROUND. The database of human Expressed Sequence Tags (dbEST) provides a potential source for identification of tissue-specific genes. This database contains sequences that originate from cDNA libraries from particular tumors, organs or cell typos. In this report, we have used the EST database to identify PRAC, a novel gene specifically expressed in human Prostate, prostate cancer, Rectum And distal Colon.METHODS. Using a computer based analysis, a cluster of sequence homologous ESTs was identified which contained ESTs derived only from human prostate cDNA libraries. The tissue specificity was examined by multiple tissue RNA dot blots and RT-PCR. The PRAC transcript and protein Mas identified using Northern blot analysis, RACE-PCR, primer extension, and western blot.RESULTS. PRAC encode a 382 nucleotide RNA found in prostate, rectum, distal colon, and in three prostate cancer cell lines; LNCaP, PC-3 and DU145. This transcript encodes a 6 kDa nuclear protein. The PRAC gene is located on chromosome 17 at position 17q21, about 4 kbp downstream from the homeodomain Hoxb-13 gene.CONCLUSIONS. Our data proves that the EST database can be a useful tool for discovery of prostate-specific genes. The nuclear localization, identification of potential phosphorylation sites, and possible cotranscription with the Hoxb-13 gene suggest that PRAC may have a regulatory role in the nucleus. Prostate 47:125-131, 2001. (C) 2001 Wiley-Liss, Inc.