Molecular cloning and expression of Xenopus p300/CBP.
Transcriptional coactivators act as signal committers from transcriptional regulators to basal transcriptional machineries. We isolated the cDNA for p300/CBP, one of the most important transcriptional coactivators, of Xenopus. We also report its regulated expression, and the effects of microinjection of its truncated form. Xenopus p300/CBP (Xp300) encodes a 2483 amino acid protein which is highly homologous with human p300. Northern hybridization analyses indicated that Xp300 mRNA is stored in the oocyte, and is present throughout early embryogenesis of this species. In situ hybridization studies have revealed that Xp300 mRNA localization is ubiquitous throughout early embryogenesis, but that in later stages it is predominant in the neural region. Among adult tissues, Xp300 mRNA was clearly detected in some tissues, suggesting that Xp300 functions as a transcriptional regulator in various tissues. Microinjection of a carboxy-terminal-truncated form of Xp300 RNA into both cells of Xenopus two-blastomere stage embryos invoked the malformation of the embryos. The neural plates of Xp300 RNA-injected embryos were loose and the trunk area was heavily contracted. These results suggest that Xp300 is indispensable for normal development of the early embryo, especially in neural formation.
Fujii
G
G
Hirohashi Cell Configuration Project, ERATO, JST, Tsukuba Research Consortium, 5-9-4, Tokodai, Tsukuba, Ibaraki 300-26, Japan.
Tsuchiya
R
R
Itoh
Y
Y
Tashiro
K
K
Hirohashi
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis; D019143:Evolution, Molecular; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D014981:Xenopus / Q000196:embryology / Q000235:genetics*
D018076:DNA, Complementary; D009687:Nuclear Proteins; D012333:RNA, Messenger; D015534:Trans-Activators