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Sequence and complementation analysis of recF genes from Escherichia coli, Salmonella typhimurium, Pseudomonas putida and Bacillus subtilis: evidence for an essential phosphate binding loop.
Nucleic Acids Res. 1992 Feb 25; 20(4):839-45.NA

Abstract

We have compared the recF genes from Escherichia coli K-12, Salmonella typhimurium, Pseudomonas putida, and Bacillus subtilis at the DNA and amino acid sequence levels. To do this we determined the complete nucleotide sequence of the recF gene from Salmonella typhimurium and we completed the nucleotide sequence of recF gene from Pseudomonas putida begun by Fujita et al. (1). We found that the RecF proteins encoded by these two genes contain respectively 92% and 38% amino acid identity with the E. coli RecF protein. Additionally, we have found that the S. typhimurium and P. putida recF genes will complement an E. coli recF mutant, but the recF gene from Bacillus subtilis [showing about 20% identity with E. coli (2)] will not. Amino acid sequence alignment of the four proteins identified four highly conserved regions. Two of these regions are part of a putative phosphate binding loop. In one region (position 36), we changed the lysine codon (which is essential for ATPase, GTPase and kinase activity in other proteins having this phosphate binding loop) to an arginine codon. We then tested this mutation (recF4101) on a multicopy plasmid for its ability to complement a recF chromosomal mutation and on the E. coli chromosome for its effect on sensitivity to UV irradiation. The strain with recF4101 on its chromosome is as sensitive as a null recF mutant strain. The strain with the plasmid-borne mutant allele is however more UV resistant than the null mutant strain. We conclude that lysine-36 and possibly a phosphate binding loop is essential for full recF activity. Lastly we made two chimeric recF genes by exchanging the amino terminal 48 amino acids of the S. typhimurium and E. coli recF genes. Both chimeras could complement E. coli chromosomal recF mutations.

Authors+Show Affiliations

Department of Molecular and Cell Biology, University of California, Berkeley 94720.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

1542576

Citation

Sandler, S J., et al. "Sequence and Complementation Analysis of recF Genes From Escherichia Coli, Salmonella Typhimurium, Pseudomonas Putida and Bacillus Subtilis: Evidence for an Essential Phosphate Binding Loop." Nucleic Acids Research, vol. 20, no. 4, 1992, pp. 839-45.
Sandler SJ, Chackerian B, Li JT, et al. Sequence and complementation analysis of recF genes from Escherichia coli, Salmonella typhimurium, Pseudomonas putida and Bacillus subtilis: evidence for an essential phosphate binding loop. Nucleic Acids Res. 1992;20(4):839-45.
Sandler, S. J., Chackerian, B., Li, J. T., & Clark, A. J. (1992). Sequence and complementation analysis of recF genes from Escherichia coli, Salmonella typhimurium, Pseudomonas putida and Bacillus subtilis: evidence for an essential phosphate binding loop. Nucleic Acids Research, 20(4), 839-45.
Sandler SJ, et al. Sequence and Complementation Analysis of recF Genes From Escherichia Coli, Salmonella Typhimurium, Pseudomonas Putida and Bacillus Subtilis: Evidence for an Essential Phosphate Binding Loop. Nucleic Acids Res. 1992 Feb 25;20(4):839-45. PubMed PMID: 1542576.
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TY - JOUR T1 - Sequence and complementation analysis of recF genes from Escherichia coli, Salmonella typhimurium, Pseudomonas putida and Bacillus subtilis: evidence for an essential phosphate binding loop. AU - Sandler,S J, AU - Chackerian,B, AU - Li,J T, AU - Clark,A J, PY - 1992/2/25/pubmed PY - 1992/2/25/medline PY - 1992/2/25/entrez SP - 839 EP - 45 JF - Nucleic acids research JO - Nucleic Acids Res VL - 20 IS - 4 N2 - We have compared the recF genes from Escherichia coli K-12, Salmonella typhimurium, Pseudomonas putida, and Bacillus subtilis at the DNA and amino acid sequence levels. To do this we determined the complete nucleotide sequence of the recF gene from Salmonella typhimurium and we completed the nucleotide sequence of recF gene from Pseudomonas putida begun by Fujita et al. (1). We found that the RecF proteins encoded by these two genes contain respectively 92% and 38% amino acid identity with the E. coli RecF protein. Additionally, we have found that the S. typhimurium and P. putida recF genes will complement an E. coli recF mutant, but the recF gene from Bacillus subtilis [showing about 20% identity with E. coli (2)] will not. Amino acid sequence alignment of the four proteins identified four highly conserved regions. Two of these regions are part of a putative phosphate binding loop. In one region (position 36), we changed the lysine codon (which is essential for ATPase, GTPase and kinase activity in other proteins having this phosphate binding loop) to an arginine codon. We then tested this mutation (recF4101) on a multicopy plasmid for its ability to complement a recF chromosomal mutation and on the E. coli chromosome for its effect on sensitivity to UV irradiation. The strain with recF4101 on its chromosome is as sensitive as a null recF mutant strain. The strain with the plasmid-borne mutant allele is however more UV resistant than the null mutant strain. We conclude that lysine-36 and possibly a phosphate binding loop is essential for full recF activity. Lastly we made two chimeric recF genes by exchanging the amino terminal 48 amino acids of the S. typhimurium and E. coli recF genes. Both chimeras could complement E. coli chromosomal recF mutations. SN - 0305-1048 UR - https://www.unboundmedicine.com/medline/citation/1542576/Sequence_and_complementation_analysis_of_recF_genes_from_Escherichia_coli_Salmonella_typhimurium_Pseudomonas_putida_and_Bacillus_subtilis:_evidence_for_an_essential_phosphate_binding_loop_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/20.4.839 DB - PRIME DP - Unbound Medicine ER -