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 types. 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 was 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.
Copyright 2001 Wiley-Liss, Inc.