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Mutation analysis of the Pip interaction domain reveals critical residues for protein-protein interactions.
- Source :
-
Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 1999 Mar 16; Vol. 96 (6), pp. 2740-5. - Publication Year :
- 1999
-
Abstract
- The PU.1 interaction partner (Pip) is a member of the interferon regulatory factor family that regulates gene expression through heterodimerization with the ETS transcription factor PU.1. Binding of Pip alone to DNA is weak, and usually it is recruited by phosphorylated PU.1 to form a strong ternary complex with specific DNA sequences. An approach combining sequence homology analysis, secondary structure predictions, and a precise mutational strategy has been used to determine critical residues within the Pip heterodimerization domain that contribute to ternary complex formation. We have delimited the Pip interaction domain to residues 245-422 by using deletion analysis. Site-directed mutagenesis of conserved polar amino acids within two predicted alpha-helices contained in this region, and which are highly conserved in the IRF family, confirmed the importance of these residues for Pip-PU.1 interaction with DNA as well as for trans-activation activity. Our results suggest the existence of a functional epitope essential for heterodimerization between Pip and PU.1 and possibly, in general, between interferon regulatory factor family members and their partners.
- Subjects :
- Amino Acid Sequence
Animals
Binding Sites genetics
DNA Mutational Analysis
DNA-Binding Proteins genetics
DNA-Binding Proteins metabolism
Interferon Regulatory Factors
Molecular Sequence Data
Protein Binding
Proto-Oncogene Proteins metabolism
Trans-Activators genetics
Trans-Activators metabolism
DNA-Binding Proteins chemistry
Proto-Oncogene Proteins chemistry
Trans-Activators chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 0027-8424
- Volume :
- 96
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Publication Type :
- Academic Journal
- Accession number :
- 10077581
- Full Text :
- https://doi.org/10.1073/pnas.96.6.2740