A Dimeric Mutant of the Homotetrameric Single-Stranded DNA Binding Protein from Escherichia coli
- 17 January 2002
- journal article
- Published by Walter de Gruyter GmbH in Biological Chemistry
- Vol. 383 (9), 1325-1333
- https://doi.org/10.1515/bc.2002.151
Abstract
A single amino acid substitution (Y78R) at the dimerdimer interface of homotetrameric single stranded DNA binding protein from E.coli (EcoSSB) renders the protein a stable dimer. This dimer can bind singlestranded DNA albeit with greatly reduced affinity. In vivo this dimeric SSB cannot replace homotetrameric EcoSSB. Amino acid changes at the rim of the dimerdimer interface nearby (Q76K, Q76E) show an electrostatic interaction between a charged amino acid at position 76 and bound nucleic acid. In conclusion, nucleic acid binding to homotetrameric SSB must take place across both dimers to achieve functionally correct binding.Keywords
This publication has 34 references indexed in Scilit:
- Identification of amino acids stabilizing the tetramerization of the single stranded DNA binding protein fromEscherichia coliFEBS Letters, 1998
- Structure of the single-stranded-DNA-binding domain of replication protein A bound to DNANature, 1997
- In Vitro and in Vivo Function of the C-Terminus of Escherichia Coli Single-Stranded DNA Binding ProteinNucleic Acids Research, 1996
- Single‐stranded‐DNA‐binding proteins from human mitochondria and Escherichia coli have analogous physicochemical propertiesEuropean Journal of Biochemistry, 1994
- Multiple binding modes of the single-stranded DNA binding protein from Escherichia coli as detected by tryptophan fluorescence and site-directed mutagenesisBiochemistry, 1993
- Amino acid 55 plays a central role in tetramerization and function of Escherichia coli single‐stranded DNA binding proteinEuropean Journal of Biochemistry, 1991
- Negative co-operativity in Escherichia coli single strand binding protein-oligonucleotide interactionsJournal of Molecular Biology, 1989
- Modulation of the affinity of the single‐stranded DNA‐binding protein of Escherichia coli (E. coli SSB) to poly(dT) by site‐directed mutagenesisEuropean Journal of Biochemistry, 1989
- A general method of analysis of ligand-macromolecule equilibria using a spectroscopic signal from the ligand to monitor binding. Application to Escherichia coli single-strand binding protein-nucleic acid interactionsBiochemistry, 1987
- Tryptophan analysis of proteins in 6M guanidine hydrochloride: Modification for more general applicationAnalytical Biochemistry, 1974