Cloning and expression of the Acanthamoeba castellanii gene encoding transcription factor TFIID

• Published in: Gene Vol. 117; no. 1; pp. 91 - 97
• Main Authors: Wong, Jie-Min, Liu, Feng, Bateman, Erik
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01.08.1992; Amsterdam Elsevier; New York, NY
• Subjects: Acanthamoeba - genetics; Acanthamoeba castellanii; Amino Acid Sequence; Animals; Base Sequence; Biological and medical sciences; Blotting, Northern; Blotting, Southern; cDNA; Cloning, Molecular; Codon; DNA, Protozoan - metabolism; DNA-binding protein; Escherichia coli; Fundamental and applied biological sciences. Psychology; gene cloning; genes; Genes. Genome; keratitis; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; nucleotide sequence; Polymerase Chain Reaction; prediction; Recombinant DNA; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA polymerase II; RNA, Messenger - genetics; RNA, Protozoan - genetics; Sequence Alignment; Transcription Factor TFIID; transcription factors; Transcription Factors - genetics; Transcription Factors - metabolism; aa; A; ORF; gene cloning; SDS; D; nt; PAGE; Recombinant DNA; bp; ds; keratitis; TFIID; DNA-binding protein; PolIk; kb; IPTG; cDNA; oligo; RNA polymerase II; PCR; Protozoa; Gene; Nucleotide sequence; Rhizoflagellata; Acanthamoeba castellanii; Recombinant protein; Gene expression; Transcription factor; Biological activity; Aminoacid sequence
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(92)90494-A

We have cloned and characterized the cDNA encoding transcription factor TFIID from the eukaryote, Acanthamoeba castellanii. The gene occurs as a single species, encodes one mRNA and, presumably, a single protein. A. castellanii TFIID contains two recognizable domains, a nonconserved N-terminal domain and a highly conserved C-terminal domain. Similarities between the amino acid (aa) sequences of TFIID from several organisms are also found within the N-terminal 78 aa, suggesting a potential role in TFIID function. Full-length or truncated A. castellanii TFIID produced in Escherichia coli binds to a TATA box and is able to activate transcription in a TFIID-depleted HeLa cell extract, but the C-terminal 180-aa domain was found to be less efficient in these reactions.