Functional interactions between the MutL and Vsr proteins of Escherichia coli are dependent on the N-terminus of Vsr.
The crystal structure of the Escherichia coli Vsr endonuclease bound to a C(T/G)AGG substrate revealed that the DNA is held by a pincer composed of a trio of aromatic residues which intercalate into the major groove, and an N-terminus alpha helix which lies across the minor groove. We have constructed an N-terminus truncation (Delta14) which removes most of the alpha helix. The mutant is still fairly proficient in mediating very short patch repair. However, its endonuclease activity is considerably reduced and, in contrast to that of the wild type protein, cannot be stimulated by MutL. We had shown previously that excess Vsr in vivo causes mutagenesis, probably by inhibiting the participation of MutL in mismatch repair. The Delta14 mutant has diminished mutagenicity. In contrast, four enzymatically inactive mutants, with intact N-termini, are as mutagenic as the wild type protein. On the basis of these results we suggest that MutL causes a conformational change in the N-terminus of Vsr which enhances Vsr activity, and that this functional interaction between Vsr and MutL decreases the ability of MutL to carry out mismatch repair.
Monastiriakos
Stavroula K
SK
Department of Chemistry and Biochemistry, Concordia University, Montréal, Qué., Canada.
Doiron
Kathy M J
KM
Siponen
Marina I
MI
Cupples
Claire G
CG
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004260:DNA Repair; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000070956:MutL Proteins; D016296:Mutagenesis; D011485:Protein Binding; D011487:Protein Conformation; D017384:Sequence Deletion
D029968:Escherichia coli Proteins; C074770:MutL protein, E coli; D004706:Endodeoxyribonucleases; C071421:vsr endonuclease; D000251:Adenosine Triphosphatases; D000070956:MutL Proteins