title: cDNA cloning, DNA binding, and evolution of mammalian transcription factor IIIA.
abstract: cDNA for rat transcription factor IIIA (TFIIIA) was cloned by degenerate PCR and rapid amplification of cDNA ends. This cDNA coded for a protein with nine Cys(2)His(2) zinc fingers and a non-finger C-terminal tail; 63% amino acid (aa) sequence identity was observed with the Xenopus TFIIIA zinc finger region. Recombinant rat protein containing only the nine fingers afforded DNase I protection of the identical nucleotides protected by Xenopus laevis native TFIIIA on the Xenopus 5S RNA gene internal control region. A putative mouse TFIIIA clone was identified in an expressed sequence tag database by sequence similarity to rat TFIIIA. Recombinant nine-finger protein from this clone afforded DNase I protection of the Xenopus 5S rRNA gene like the native frog protein as did a recombinant nine-finger form of a putative human TFIIIA clone. These DNA binding results demonstrate that these clones code for the respective mammalian TFIIIAs. Rodent and human TFIIIAs share about 87% aa sequence identity in their zinc finger regions and have evolved to about the same extent as X. laevis and Xenopus borealis TFIIIAs. A monoclonal antibody against human p53 tumor suppressor bound to rat and mouse TFIIIA but not to human TFIIIA in Western blots. The N-terminal regions of rodent and human TFIIIA do not contain the oocyte-specific initiating Met and accompanying conserved residues found in fish and amphibian TFIIIAs. In their non-finger C-terminal tails, mammalian and amphibian TFIIIAs share a conserved transcription activation domain as well as conserved nuclear localization and nuclear export signals.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma College of Medicine, Oklahoma City, OK 73104, USA. jay-hanas@ouhsc.edu
authors.forename: Jay S
authors.initials: JS
authors.lastname: Hanas
authors.forename: James R
authors.initials: JR
authors.lastname: Hocker
authors.forename: Yong-Gang
authors.initials: YG
authors.lastname: Cheng
authors.forename: Megan R
authors.initials: MR
authors.lastname: Lerner
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Brackett
authors.forename: Stan A
authors.initials: SA
authors.lastname: Lightfoot
authors.forename: Rushie J
authors.initials: RJ
authors.lastname: Hanas
authors.forename: Kunapuli T
authors.initials: KT
authors.lastname: Madhusudhan
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Moreland
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D019143:Evolution, Molecular; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D051381:Rats; D011920:Rats, Inbred WF; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors