Changing the recognition site of a conjugative relaxase by rational design

Biotechnology Journal

Volumn 4, Issue 4, 2009, Pages 554-557

  González Pérez, Blanca ^a   Carballeira, José Daniel ^a   Moncalián, Gabriel ^a   de la Cruz, Fernando ^a  

^a UNIVERSITY OF CANTABRIA   (Spain)

Author keywords

Bacterial conjugation; Rational design; Relaxase

Indexed keywords

BACTERIAL CONJUGATION; DONOR CELLS; IN-VIVO; PROOF OF PRINCIPLES; RATIONAL DESIGN; RECOGNITION SITES; RELAXASE; SATURATION MUTAGENESIS; SELECTION SYSTEMS; SIDE CHAINS; SIMPLE MODELS; SPECIFIC SEQUENCES; SUBSTRATE SPECIFICITIES; WILD TYPES;

AMINES; AMINO ACIDS; BACTERIOLOGY; DESIGN; DNA; GENES; ORGANIC ACIDS;

NUCLEIC ACIDS;

BACTERIAL ENZYME; PROTEIN TRWC; RELAXASE; UNCLASSIFIED DRUG;

AMINO ACID SEQUENCE; ARTICLE; BACTERIUM CONJUGATION; DNA PROTEIN COMPLEX; DNA SEQUENCE; ENZYME SPECIFICITY; GENE LIBRARY; GENETIC SELECTION; GENETIC VARIABILITY; MOLECULAR RECOGNITION; MUTATION; NUCLEIC ACID BASE SUBSTITUTION; PRIORITY JOURNAL;

ALANINE; AMINO ACID SUBSTITUTION; BASE SEQUENCE; BINDING SITES; CONJUGATION, GENETIC; CYSTEINE; DNA NUCLEOTIDYLTRANSFERASES; DNA, BACTERIAL; ESCHERICHIA COLI; ESCHERICHIA COLI PROTEINS; MODELS, GENETIC; MODELS, MOLECULAR; MOLECULAR SEQUENCE DATA; MOLECULAR STRUCTURE; MUTAGENESIS, SITE-DIRECTED; PLASMIDS; PROTEIN BINDING; PROTEIN ENGINEERING; SUBSTRATE SPECIFICITY; VALINE;

BACTERIA (MICROORGANISMS);

EID: 65549113871     PISSN: 18606768     EISSN: 18607314     Source Type: Journal    
DOI: 10.1002/biot.200800184     Document Type: Article

References (15)

1. Gonzalez-Perez, B., Lucas, M., Cooke, L. A., Vyle, J. S. et al., Analysis of DNA processing reactions in bacterial conjugation by using suicide oligonucleotides. EMBO J. 2007, 26, 3847-3857.
2. Garcillan-Barcia, M. P., Jurado, P., Gonzalez-Perez, B., Moncalian, G. et al., Conjugative transfer can be inhibited by blocking relaxase activity within recipient cells with intrabodies. Mol. Microbiol. 2007, 63, 404-416.
3. Llosa, M., de la Cruz, F., Bacterial conjugation: A potential tool for genomic engineering. Res. Microbiol. 2005, 156, 1-6.
4. Mathias, J. R., Zhong, H., Jin, Y., Vershon, A. K., Altering the DNA-binding specificity of the yeast Mata2 homeodomain protein. J. Biol. Chem. 2001, 276, 32696-32703.
5. Namgoong, S.Y., Sankaralingam, S., Harshey R. M., Altering the DNA-binding specificity of Mu transposase in vitro. Nucleic Acids Res. 1998, 26, 3521-3527.
6. Hekman, K., Guja, K., Larkin, C., Schildbach, J. F., An intrastrand three-DNA-base interaction is a key specificity determinant of F transfer initiation and of F TraI relaxase DNA recognition and cleavage. Nucleic Acids Res. 2008, 36, 4565-4572.
7. Caryl, J.A., Thomas, C.D., Investigating the basis of substrate recognition in the pC221 relaxosome. Mol. Microbiol. 2006, 60, 1302-1318.
8. Harley, M. J., Schildbach, J. F., Swapping single-stranded DNA sequence specificities of relaxases from conjugative plasmids F and R100. Proc. Natl. Acad. Sci. 2003, 30, 11243-11248.
9. Boer, R., Russi, S., Guasch, A., Lucas, M. et al., Unveiling the molecular mechanism of a conjugative relaxase:The structure of TrwC complexed with a 27-mer DNA comprising the recognition hairpin and the cleavage site. J. Mol. Biol. 2006, 358, 857-869.
10. Guasch, A., Lucas, M., Moncalian, G., Cabezas, M. et al., Recognition and processing of the origin of transfer DNA by conjugative relaxase TrwC. Nat. Struct. Biol. 2003, 10, 1002-1010.
11. Hogrefe, H. H., Cline, J., Youngblood, G. L., Allen R. M., Creating randomized amino acid libraries with the QuikChange Multi Site-Directed Mutagenesis Kit. BioTechniques 2002, 33, 1158-1165.
12. Reetz, M.T., Kahakeaw D., Lohmer R., Addressing the numbers problem in directed evolution. ChemBioChem 2008, 9, 1797-1804.
13. Llosa, M., Bolland, S., Grandoso, G., de la Cruz, F., Conjugation-independent, site-specific recombination at the oriT of the IncW plasmid R388 mediated by TrwC. J. Bacteriol. 1994, 176, 3210-3217.
14. Bradley, D. E., Taylor, D. E., Cohen, D. R., Specification of surface mating systems among conjugative drug resistance plasmids in Escherichia coli K-12. J. Bacteriol. 1980, 143, 1466-1470.
15. de la Cruz, F., Grinsted, J., Genetic and molecular characterization of Tn21, a multiple resistance transposón from R100-1. J. Bacteriol. 1982, 151, 222-228.