CLONING AND EXPRESSION OF HUMAN NF-YC

The CCAAT box is an important element in eukaryotic promoters and NF-Y (CBF) is a conserved heterotrimeric protein binding to it. Two subuni ts, NF-YB and NF-YC, contain a histone-like motif. We cloned the compl ete cDNA coding for the human NF-YC gene. The ORF codes for a 335 aa p rotein that shows virtual identity to the rat sequence, confirming the stunning invariance of NF-Y genes across species. We expressed and pu rified the yeast homology domain of NF-YC in bacteria and performed EM SA together with the corresponding conserved domains of NF-YA and NF-Y B, obtaining a CCAAT-binding mini-NF-Y. We evaluated the expression of NF-YC and found that mRNA levels are similar in different human tissu es except in testis. (C) 1997 Elsevier Science B.V.