Cloning and expression of human NF-YC

• Published in: Gene 1997-07, Vol.193 (1), p.119-125
• Main Authors: Bellorini, Marianna, Zemzoumi, Khalid, Farina, Andrea, Berthelsen, Jens, Piaggio, Giulia, Mantovani, Roberto
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Base Sequence; Binding Sites; Biological Evolution; CCAAT; CCAAT-Enhancer-Binding Proteins; Cloning, Molecular; Conserved Sequence; DNA, Complementary - genetics; DNA-Binding Proteins - genetics; DNA-Binding Proteins - isolation & purification; DNA-Binding Proteins - metabolism; Escherichia coli - genetics; Histone fold motif; Humans; Molecular Sequence Data; NF-Y; Peptide Fragments - genetics; Peptide Fragments - isolation & purification; Peptide Fragments - metabolism; Protein Binding; Protein Conformation; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; RNA, Messenger - genetics; Sequence Analysis, DNA; Species Specificity; Tissue Distribution; Transcription
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/S0378-1119(97)00109-1

The CCAAT box is an important element in eukaryotic promoters and NF-Y (CBF) is a conserved heterotrimeric protein binding to it. Two subunits, NF-YB and NF-YC, contain a histone-like motif. We cloned the complete cDNA coding for the human NF-YC gene. The ORF codes for a 335 aa protein that shows virtual identity to the rat sequence, confirming the stunning invariance of NF-Y genes across species. We expressed and purified the yeast homology domain of NF-YC in bacteria and performed EMSA together with the corresponding conserved domains of NF-YA and NF-YB, obtaining a CCAAT-binding mini-NF-Y. We evaluated the expression of NF-YC and found that mRNA levels are similar in different human tissues except in testis.