Identification of N-terminal minimal transactivation domain of CBP, p300 and caenorhabditis elegans homologues.
CBP/p300 is a multidomain transcriptional cofactor that acts in junction with other factors to regulate transcription. To elucidate the domain function of CBP, we fused its dissected fragments to Ga14 DNA-binding domain and transfected the deletion mutants into several cell lines. First, we found that the minimal transactivation domain (MTD) at the N-terminal portion maps to between 344 and 451 aa, and shows activity in a cell-type dependent manner. Second, we cloned C. elegans homologues corresponding to the MTD by RT-PCR and identified the three related products, two of which exhibited weak transcriptional activity. Finally, by means of the yeast two hybrid screening using MTD as a bait, we cloned hypoxia-inducible factor (HIF) 1 alpha and Stat2 cDNAs. These results suggested a functional role of MTD located at the N-terminal region of CBP/p300 in connecting to transcriptional factors.
Yoshida
E
E
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
Nakajima
T
T
Murakami
K
K
Fukamizu
A
A
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D050882:CREB-Binding Protein; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D005091:Exons; D017238:Genes, Helminth; D015801:Helminth Proteins / Q000096:biosynthesis / Q000737:chemistry; D007438:Introns; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017384:Sequence Deletion; D015534:Trans-Activators; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*; D015533:Transcriptional Activation; D014162:Transfection
D015801:Helminth Proteins; D009687:Nuclear Proteins; D011994:Recombinant Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D050882:CREB-Binding Protein