Cloning And Identification Of Human FAM33A Promoter

FAM33A (family with sequence similarity 33), also known as Ska2 (spindle and KT associated 2), is a recently identified molecule involved in cell proliferation and cell cycle regulation. PRR11 (proline rich 11) was initially being identified from human full-length cDNA in 2004, which is a proline-rich encoded gene located in chromosome 17. To further investigate its transcriptional regulatory mechanism, PRR11 and FAM33A promoter has been cloned and identified in the present study for the first time.At first, the transcriptional start sites for FAM33A gene have been identified by 5' RACE (rapid amplification of cDNA ends) and bioinformatics analysis. Furthermore, several overlapping genomic fragments from the 5'- flanking region of PRR11 and FAM33A gene have been cloned into pGL3-basic vector to construct PRR11 and FAM33A promoter reporters. Luciferase reporter assay indicated that FAM33A promoter region is mainly located in a 590 bp region upstream of the transcriptional start site. The overlapping region shared by PRR11 and FAM33A has promoter activity to some degree, thus this overlapping region may serve as co-modulator both of PRR11 and FAM33A promoter. Transcriptional factor binding analysis indicated that, FAM33A promoter contains GC box, as well as other putative transcriptional factor binding sites such as Sp1, E2F, and GATA-1. These results suggested that transcriptional factors such as Sp1, E2F, and GATA-1 might be involved in the transcriptional regulation of FAM33A gene.