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PMID: 7739566 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

DNA-binding and transactivation properties of Pax-6: three amino acids in the paired domain are responsible for the different sequence recognition of Pax-6 and BSAP (Pax-5).

Molecular and cellular biology ·Vol. 15 ·No. 5 ·1995-05-00 ·Pages 2858-71

Czerny T, Busslinger M

Abstract

Pax-6 is known to be a key regulator of vertebrate eye development. We have now isolated cDNA for an invertebrate Pax-6 protein from sea urchin embryos. Transcripts of this gene first appear during development at the gastrula stage and are later expressed at high levels in the tube foot of the adult sea urchin. The sea urchin Pax-6 protein is highly homologous throughout the whole protein to its vertebrate counterpart with the paired domain and homeodomain being virtually identical. Consequently, we found that the DNA-binding and transactivation properties of the sea urchin and mouse Pax-6 proteins are very similar, if not identical. A potent activation domain capable of stimulating transcription from proximal promoter and distal enhancer positions was localized within the C-terminal sequences of both the sea urchin and mouse Pax-6 proteins. The homeodomain of Pax-6 was shown to cooperatively dimerize on DNA sequences consisting of an inverted repeat of the TAAT motif with a preferred spacing of 3 nucleotides. The consensus recognition sequence of the Pax-6 paired domain deviates primarily only at one position from that of BSAP (Pax-5), and yet the two proteins exhibit largely different binding specificities for individual, naturally occurring sites. By creating Pax-6-BSAP fusion proteins, we were able to identify a short amino acid stretch in the N-terminal part of the paired domain which is responsible for these differences in DNA-binding specificity. Mutation of three Pax-6-specific residues in this region (at positions 42, 44, and 47 of the paired domain) to the corresponding amino acids of BSAP resulted in a complete switch of the DNA-binding specificity from Pax-6 to BSAP. These three amino acids were furthermore shown to discriminate between the Pax-6- and BSAP-specific nucleotide at the divergent position of the two consensus recognition sequences.

Related Genes
Pax-6
MeSH Terms
Amino Acid Sequence Animals Base Sequence Binding Sites/genetics Cloning, Molecular Consensus Sequence DNA Primers/genetics DNA, Complementary/genetics DNA-Binding Proteins/genetics,metabolism Eye Proteins Gene Expression Regulation, Developmental Homeodomain Proteins Mice Molecular Sequence Data Nuclear Proteins/genetics,metabolism PAX5 Transcription Factor PAX6 Transcription Factor Paired Box Transcription Factors Repressor Proteins Sea Urchins/genetics Sequence Homology, Amino Acid Species Specificity Trans-Activators/genetics,metabolism Transcription Factors
Chemicals
DNA Primers DNA, Complementary DNA-Binding Proteins Eye Proteins Homeodomain Proteins Nuclear Proteins PAX5 Transcription Factor PAX6 Transcription Factor Paired Box Transcription Factors Pax5 protein, mouse Pax6 protein, mouse Repressor Proteins Trans-Activators Transcription Factors
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Czerny T
Research Institute of Molecular Pathology, Vienna, Austria.
Busslinger M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.15.5.2858
Published
1995-05-00
Pages
2858-71
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC230517
Subset
IM
Databases
GENBANK
U14621
External Links
PubMed source DOI: 10.1128/MCB.15.5.2858 PMC Full Text View journal details
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