FimY of Salmonella enterica serovar Typhimurium functions as a DNA-binding protein and binds the fimZ promoter

Ke Chuan Wang, Yuan Hsun Hsu, Yi Ning Huang, Jiunn Horng Lin, Kuang Sheng Yeh

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Salmonella enterica serovar Typhimurium produces type 1 fimbriae with binding specificity to mannose residues. Elements involved in fimbrial structural biosynthesis, transport, and regulation are encoded by the fim gene cluster. FimZ, FimY, FimW, STM0551, and an arginine transfer RNA (fimU) were previously demonstrated to regulate fimbrial expression. The amino acid sequences of the C-terminal portion of FimY revealed similarity with those of LuxR-like proteins. Electrophoretic mobility shift assays indicated that FimY possessed DNA-binding capacity and bound a 605-bp DNA fragment spanning the intergenic region between fimY and fimZ, while a FimY protein harboring a double mutation in the C-terminal helix-turn-helix region containing a glycine (G) to aspartate (D) substitution at residue 189 and isoleucine (I) to lysine (K) substitution at residue 195 lost its ability to bind this DNA fragment. A lux box sequence (5'-TCTGTTATTACATAACAAATACT-3') within the fimZ promoter was required for binding. None of the DNA fragments derived from the promoters for fimA, fimY, or fimW was shifted by FimY. Pull-down assays showed that there were physical protein/protein interactions between FimY and FimZ. We propose that in the regulatory circuit of type 1 fimbriae, FimY functions as a DNA-binding protein to activate fimZ, and a FimY-FimZ protein complex may form to regulate other fim genes. Confirming these proposals requires further study.

Original languageEnglish
Pages (from-to)496-503
Number of pages8
JournalMicrobiological Research
Volume169
Issue number7-8
DOIs
Publication statusPublished - 2014

Fingerprint

Salmonella enterica
DNA-Binding Proteins
DNA
Proteins
Intergenic DNA
Isoleucine
Electrophoretic Mobility Shift Assay
Mannose
Multigene Family
Transfer RNA
Aspartic Acid
Glycine
Lysine
Arginine
Amino Acid Sequence
Serogroup
Mutation
Genes

Keywords

  • DNA binding protein
  • Salmonella
  • Type 1 fimbriae

ASJC Scopus subject areas

  • Microbiology
  • Medicine(all)

Cite this

FimY of Salmonella enterica serovar Typhimurium functions as a DNA-binding protein and binds the fimZ promoter. / Wang, Ke Chuan; Hsu, Yuan Hsun; Huang, Yi Ning; Lin, Jiunn Horng; Yeh, Kuang Sheng.

In: Microbiological Research, Vol. 169, No. 7-8, 2014, p. 496-503.

Research output: Contribution to journalArticle

Wang, Ke Chuan ; Hsu, Yuan Hsun ; Huang, Yi Ning ; Lin, Jiunn Horng ; Yeh, Kuang Sheng. / FimY of Salmonella enterica serovar Typhimurium functions as a DNA-binding protein and binds the fimZ promoter. In: Microbiological Research. 2014 ; Vol. 169, No. 7-8. pp. 496-503.
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AU - Lin, Jiunn Horng

AU - Yeh, Kuang Sheng

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N2 - Salmonella enterica serovar Typhimurium produces type 1 fimbriae with binding specificity to mannose residues. Elements involved in fimbrial structural biosynthesis, transport, and regulation are encoded by the fim gene cluster. FimZ, FimY, FimW, STM0551, and an arginine transfer RNA (fimU) were previously demonstrated to regulate fimbrial expression. The amino acid sequences of the C-terminal portion of FimY revealed similarity with those of LuxR-like proteins. Electrophoretic mobility shift assays indicated that FimY possessed DNA-binding capacity and bound a 605-bp DNA fragment spanning the intergenic region between fimY and fimZ, while a FimY protein harboring a double mutation in the C-terminal helix-turn-helix region containing a glycine (G) to aspartate (D) substitution at residue 189 and isoleucine (I) to lysine (K) substitution at residue 195 lost its ability to bind this DNA fragment. A lux box sequence (5'-TCTGTTATTACATAACAAATACT-3') within the fimZ promoter was required for binding. None of the DNA fragments derived from the promoters for fimA, fimY, or fimW was shifted by FimY. Pull-down assays showed that there were physical protein/protein interactions between FimY and FimZ. We propose that in the regulatory circuit of type 1 fimbriae, FimY functions as a DNA-binding protein to activate fimZ, and a FimY-FimZ protein complex may form to regulate other fim genes. Confirming these proposals requires further study.

AB - Salmonella enterica serovar Typhimurium produces type 1 fimbriae with binding specificity to mannose residues. Elements involved in fimbrial structural biosynthesis, transport, and regulation are encoded by the fim gene cluster. FimZ, FimY, FimW, STM0551, and an arginine transfer RNA (fimU) were previously demonstrated to regulate fimbrial expression. The amino acid sequences of the C-terminal portion of FimY revealed similarity with those of LuxR-like proteins. Electrophoretic mobility shift assays indicated that FimY possessed DNA-binding capacity and bound a 605-bp DNA fragment spanning the intergenic region between fimY and fimZ, while a FimY protein harboring a double mutation in the C-terminal helix-turn-helix region containing a glycine (G) to aspartate (D) substitution at residue 189 and isoleucine (I) to lysine (K) substitution at residue 195 lost its ability to bind this DNA fragment. A lux box sequence (5'-TCTGTTATTACATAACAAATACT-3') within the fimZ promoter was required for binding. None of the DNA fragments derived from the promoters for fimA, fimY, or fimW was shifted by FimY. Pull-down assays showed that there were physical protein/protein interactions between FimY and FimZ. We propose that in the regulatory circuit of type 1 fimbriae, FimY functions as a DNA-binding protein to activate fimZ, and a FimY-FimZ protein complex may form to regulate other fim genes. Confirming these proposals requires further study.

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