cDNA cloning, DNA binding, and evolution of mammalian transcription factor IIIA

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 finge...

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Bibliographic Details
Published in:Gene 2002-01, Vol.282 (1-2), p.43-52
Main Authors: Hanas, Jay S, Hocker, James R, Cheng, Yong-Gang, Lerner, Megan R, Brackett, Daniel J, Lightfoot, Stan A, Hanas, Rushie J, Madhusudhan, Kunapuli T, Moreland, Rodney J
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Cloning, Molecular
DNA, Complementary - chemistry
DNA, Complementary - genetics
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Evolution, Molecular
Gene Expression
Humans
Mice
Molecular Sequence Data
Protein Binding
Rats
Rats, Inbred WF
Rats, Sprague-Dawley
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Sequence Alignment
Sequence Analysis, DNA
Sequence Homology, Amino Acid
TFIIA protein
Transcription Factor TFIIIA
Transcription Factors - genetics
Transcription Factors - metabolism
Xenopus borealis
Xenopus laevis
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Summary: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.
ISSN:0378-1119
1879-0038
DOI:10.1016/s0378-1119(01)00796-x