Cloning and characterisation of the RBCC728/TRIM36 zinc-binding protein from the tumor suppressor gene region at chromosome 5q22.3.

DNA alterations at chromosome 5q22 occur frequently in different types of tumors including urological cancers. Previously, we narrowed a common target region between loci D5S659 and D5S2055 at chromosome 5q22.3 by microsatellite allelotyping. After constructing a BAC contig and shot-gun sequencing we identified a putative exon by the NIX software package. By PCR cloning using the putative exon (5qex5) specific primers and primers directed to the vector (lambdaZAPII) sequence of human brain and kidney cDNA libraries, we obtained a full-length cDNA of 4074 bp of the new gene RBCC728/TRIM36 (GenBank accession no.) with an ORF coding for a protein of 728 amino acids. The TRIM36, which is a new member of the tripartite motif (TRIM) gene family, shows a RING finger C3HC4 structure, two B-box, a coiled-coil, a fibronectin type III and a C-terminal domain of unknown function (SPRY). The TRIM36 has a weak homology to MID1. Immunohistochemistry of recombinant and native TRIM36 displays a cytoplasmic, slightly filamentous staining pattern in COS-7 cells. The TRIM36 is expressed in adult testis, brain, prostate, kidney, heart and lung. A variable level of TRIM36 expression was detected by Q-RT-PCR in conventional RCC, while the gene was consequently upregulated in PCs. We did not find mutation in the open reading frame of the TRIM36 in cancer cells. The overexpression of the TRIM36 in the vast majority of prostate cancer suggest that this gene might be involved in the prostate tumorigenesis.