Cloning And Functional Analysis Of Human Heart Developmental Candidate Gene ZNF649

Cardiac differentiation involves a cascade of coordinated gene expression that regulates cell proliferation and matrix protein formation in a defined temporo-spatial manner. Many of the KRAB-ZFPs are involved in cardiac development or cardiovascular diseases. Here we report the identification and characterization of a novel human zinc-finger gene named ZNF649. The cDNA of ZNF649 is 3176 bp, encoding a protein of 505 amino acids in the nuclei with an N-terminal KRAB domain and 10 classical zinc finger C2H2 motifs in the C-terminus. Scanning with bioinformatics methods, ZNF649 is identical on chromosome 19ql3.41, spanning approximately 17 Kb on the genome and organized into 5 exons.Northern blot analysis indicates that ZNF649 is expressed in most of the examined human adult and embryonic tissues, at a higher level in heart and skeletal muscle. ZNF649 is a transcription suppressor when fused to Gal-4 DNA-binding domain and cotransfected with VP-16. Overexpression of ZNF649 in COS-7 cells inhibits the transcriptional activities of SRE and AP-1 which are downstream effectors of MAPK pathway. Deletion analysis with a series of truncated fusion proteins indicates that the KRAB motif is a basal repression domain when the truncated fusion proteins were assayed for the transcriptional activities of...