Neisseria gonorrhoeae FitA interacts with FitB to bind DNA through its ribbon-helix-helix motif

J. Scott Wilbur; Peter T. Chivers; Kirsten Mattison; Laura Potter; Richard G. Brennan; Magdalene So

doi:10.1021/bi0511080

Neisseria gonorrhoeae FitA interacts with FitB to bind DNA through its ribbon-helix-helix motif

J. Scott Wilbur, Peter T. Chivers, Kirsten Mattison, Laura Potter, Richard G. Brennan, Magdalene So

Research output: Contribution to journal › Article › peer-review

31 Scopus citations

Abstract

The fit locus, encoding two proteins, FitA and FitB, was identified in a genetic screen for Neisseria gonorrhoeae determinants that affect trafficking across polarized epithelial cells. To better understand how the locus may control these activities, we have undertaken a biochemical analysis of FitA and FitB. FitA is a DNA-binding protein with a putative ribbon-helix-helix (RHH) motif. Purified FitA forms a homodimer that binds a 150 bp fit promoter sequence containing the translational start site. A putative β strand mutant of FitA, FitA(R7A), is unable to bind this DNA, supporting further that FitA is a RHH protein. FitB interacts with FitA to form a 98 kDa complex. FitA/B binds DNA with a 38-fold higher affinity than the FitA homodimer. In DNase I footprint assays, FitA/B protects a 62-bp region within the fit promoter containing the predicted -10 sequence and an 8-bp inverted repeat, TGCTATCA-N ₁₂-TGATAGCA. FitA/B_HiS is able to bind to either half-site alone with high affinity.

Original language	English (US)
Pages (from-to)	12515-12524
Number of pages	10
Journal	Biochemistry
Volume	44
Issue number	37
DOIs	https://doi.org/10.1021/bi0511080
State	Published - Sep 20 2005
Externally published	Yes

ASJC Scopus subject areas

Biochemistry

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title = "Neisseria gonorrhoeae FitA interacts with FitB to bind DNA through its ribbon-helix-helix motif",

abstract = "The fit locus, encoding two proteins, FitA and FitB, was identified in a genetic screen for Neisseria gonorrhoeae determinants that affect trafficking across polarized epithelial cells. To better understand how the locus may control these activities, we have undertaken a biochemical analysis of FitA and FitB. FitA is a DNA-binding protein with a putative ribbon-helix-helix (RHH) motif. Purified FitA forms a homodimer that binds a 150 bp fit promoter sequence containing the translational start site. A putative β strand mutant of FitA, FitA(R7A), is unable to bind this DNA, supporting further that FitA is a RHH protein. FitB interacts with FitA to form a 98 kDa complex. FitA/B binds DNA with a 38-fold higher affinity than the FitA homodimer. In DNase I footprint assays, FitA/B protects a 62-bp region within the fit promoter containing the predicted -10 sequence and an 8-bp inverted repeat, TGCTATCA-N 12-TGATAGCA. FitA/BHiS is able to bind to either half-site alone with high affinity.",

author = "Wilbur, {J. Scott} and Chivers, {Peter T.} and Kirsten Mattison and Laura Potter and Brennan, {Richard G.} and Magdalene So",

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AU - Wilbur, J. Scott

AU - Chivers, Peter T.

AU - Mattison, Kirsten

AU - Potter, Laura

AU - Brennan, Richard G.

AU - So, Magdalene

PY - 2005/9/20

Y1 - 2005/9/20

N2 - The fit locus, encoding two proteins, FitA and FitB, was identified in a genetic screen for Neisseria gonorrhoeae determinants that affect trafficking across polarized epithelial cells. To better understand how the locus may control these activities, we have undertaken a biochemical analysis of FitA and FitB. FitA is a DNA-binding protein with a putative ribbon-helix-helix (RHH) motif. Purified FitA forms a homodimer that binds a 150 bp fit promoter sequence containing the translational start site. A putative β strand mutant of FitA, FitA(R7A), is unable to bind this DNA, supporting further that FitA is a RHH protein. FitB interacts with FitA to form a 98 kDa complex. FitA/B binds DNA with a 38-fold higher affinity than the FitA homodimer. In DNase I footprint assays, FitA/B protects a 62-bp region within the fit promoter containing the predicted -10 sequence and an 8-bp inverted repeat, TGCTATCA-N 12-TGATAGCA. FitA/BHiS is able to bind to either half-site alone with high affinity.

AB - The fit locus, encoding two proteins, FitA and FitB, was identified in a genetic screen for Neisseria gonorrhoeae determinants that affect trafficking across polarized epithelial cells. To better understand how the locus may control these activities, we have undertaken a biochemical analysis of FitA and FitB. FitA is a DNA-binding protein with a putative ribbon-helix-helix (RHH) motif. Purified FitA forms a homodimer that binds a 150 bp fit promoter sequence containing the translational start site. A putative β strand mutant of FitA, FitA(R7A), is unable to bind this DNA, supporting further that FitA is a RHH protein. FitB interacts with FitA to form a 98 kDa complex. FitA/B binds DNA with a 38-fold higher affinity than the FitA homodimer. In DNase I footprint assays, FitA/B protects a 62-bp region within the fit promoter containing the predicted -10 sequence and an 8-bp inverted repeat, TGCTATCA-N 12-TGATAGCA. FitA/BHiS is able to bind to either half-site alone with high affinity.

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Neisseria gonorrhoeae FitA interacts with FitB to bind DNA through its ribbon-helix-helix motif

Abstract

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