H-NS Oligomerization Domain Structure Reveals the Mechanism for High Order Self-association of the Intact Protein

Esposito, Diego; Petrovic, Arsen; Harris, Richard; Ono, Shusuke; Eccleston, John F.; Mbabaali, A.; Haq, Ihtshamul; Higgins, Christopher F.; Hinton, Jay C. D.; Driscoll, Paul C.; Ladbury, John E.

doi:10.1016/s0022-2836(02)01141-5

Cited by 124 publications

(192 citation statements)

References 29 publications

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“…Recent studies have established that H-NS is assembled as a polymeric superhelical scaffold (around which DNA is wrapped) through two oligomerization motifs that are referred to as site 1 and site 2 interfaces, respectively (3,5,8,22,54,57,58). The former involves the N-terminal domain of H-NS (residue 2 to approximately residue 47) and the latter the intermediate or linker domain (residues 58 to 84), and polymerization is achieved by the alternation of site 1-site 1 (headto-head) and site 2-site 2 (tail-to-tail) interactions between the adjacent protomers in the scaffold (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, that the other dominant-negative H-NS variants, such as ⌬93, Y97C, P116S, and I119T, which are unaffected for transcriptional polarity relief, are proficient for oligomerization through both interfaces (Fig. 8E and F) (3,5,8,22,54,57).…”

Section: Discussionmentioning

confidence: 99%

“…(Amino acid residues in H-NS are numbered here beginning from the N-terminal methionine as 1 [65], even though this residue is posttranslationally removed from the mature protein.) The presence of the two dimerization interfaces allows H-NS to assemble (either alone or with its paralogous partner StpA [53,55,60,65,67]) into a helical polymeric scaffold around which DNA is bound (3,22). Binding of H-NS to DNA results in several structural (including supercoiling) alterations that have been variously referred to as bending, bridging, coating, looping, and stiffening of the DNA (16, 20,…”

mentioning

confidence: 99%

“…YdgT (also called Cnu [30]) and Hha belong to the H-NS family of proteins (33), whose prototypic member H-NS participates in both regulation of transcription initiation and nucleoid structure (reviewed in references 16, 20, 23, and 59). Genetic, biophysical, and structural studies have shown that the 137-amino acid H-NS protein possesses two separate dimerization interfaces, between approximately residues 2 and 47 and residues 58 and 84, respectively, and a third C-terminal DNA-binding domain between residues 92 and 137 (3,5,8,22,54,57,58). (Amino acid residues in H-NS are numbered here beginning from the N-terminal methionine as 1 [65], even though this residue is posttranslationally removed from the mature protein.)…”

mentioning

confidence: 99%

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Modulation of Rho-Dependent Transcription Termination in Escherichia coli by the H-NS Family of Proteins

Saxena

Gowrishankar

2011

J Bacteriol

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Nascent transcripts in Escherichia coli that fail to be simultaneously translated are subject to a factordependent mechanism of termination (also termed a polarity) that involves the proteins Rho and NusG. In this study, we found that overexpression of YdgT suppressed the polarity relief phenotypes and restored the efficiency of termination in rho or nusG mutants. YdgT and Hha belong to the H-NS and StpA family of proteins that repress a large number of genes in Gram-negative bacteria. Variants of H-NS defective in one or the other of its two dimerization domains, but not those defective in DNA binding alone, also conferred a similar suppression phenotype in rho and nusG mutants. YdgT overexpression was associated with derepression of proU, a prototypical H-NS-silenced locus. Polarity relief conferred by rho or nusG was unaffected in a derivative completely deficient for both H-NS and StpA, although the suppression effects of YdgT or the oligomerizationdefective H-NS variants were abolished in this background. Transcription elongation rates in vivo were unaffected in any of the suppressor-bearing strains. Finally, the polarity defects of rho and nusG mutants were exacerbated by Hha and YdgT deficiency. A model is proposed that invokes a novel role for the polymeric architectural scaffold formed on DNA by H-NS and StpA independent of the gene-silencing functions of these nucleoid proteins, in modulating Rho-dependent transcription termination such that interruption of the scaffold (as obtained by expression either of the H-NS oligomerization variants or of YdgT) is associated with improved termination efficiency in the rho and nusG mutants.Translation is a cotranscriptional process in both eubacteria and archaebacteria (1,9,25,45,50), and it has been proposed that such coupling is a defining characteristic of prokaryotic life (34, 64). The maintenance of transcription-translation coupling in a prokaryotic cell would require dynamic inter-regulation between the binding and progression of a pioneer ribosome on the nascent transcript on the one hand and the rate of transcription elongation on the other (9, 45); however, the detailed mechanisms by which such regulation is achieved are not known. Furthermore, in bacteria such as Escherichia coli, nascent transcripts that are not simultaneously translated are subject to a mechanism of factor-dependent transcription termination (also referred to as transcriptional polarity) (reviewed in references 1, 6, 15, 40, 44, 47, and 52), so that the occurrence of translation-uncoupled transcription is minimized within the cells (11, 43).Factor-dependent (also called Rho-dependent) transcription termination is mediated by, among others, the Rho and NusG proteins (1,6,15,39,40,44,47,52). Although the structures of the two proteins have been determined and several of their biochemical properties have been characterized, the precise mechanisms of their action in transcription termination remain unclear, even controversial. Rho is an RNAbinding protein and possesses RNA-dependent ATPa...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Modulation of Rho-Dependent Transcription Termination in Escherichia coli by the H-NS Family of Proteins

Saxena

Gowrishankar

2011

J Bacteriol

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“…The N-terminal domain forms dimers in solution [19,24]. We had previously identified a Hha binding region in the N-terminal domain of E. coli H-NS and demonstrated that Arg12 is essential for Hha binding [17].…”

Section: Hha Binds Exclusively To the N-terminal Domain Of Ech-ns Andmentioning

confidence: 99%

N9L and L9N mutations toggle Hha binding and hemolysin regulation by Escherichia coli and Vibrio cholerae H‐NS

et al. 2009

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a b s t r a c tProteins of the Hha/YmoA family co-regulate with H-NS the expression of virulence factors in Enterobacteriaceae. Vibrio cholerae lacks Hha-like proteins and its H-NS (vcH-NS) is unable to bind Hha, in spite of the conservation of a key residue for Hha binding by Escherichia coli H-NS (ecH-NS). Exchange of the residues in position 9 between vcH-NS and ecH-NS strongly reduces Hha binding by ecH-NS and introduces it in vcH-NS. These mutations strongly affect the repression of the hemolysin operon in E. coli and the electrophoretic mobility of complexes formed with a DNA fragment containing its regulatory region.

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Structural similarities and differences in H‐NS family proteins revealed by the N‐terminal structure of TurB in Pseudomonas putida KT2440

et al. 2016

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H-NS family proteins play key roles in bacterial nucleoid compaction and global transcription. MvaT homologues in Pseudomonas have almost negligible amino acid sequence identity with H-NS, but can complement an hns-related phenotype of Escherichia coli. Here, we report the crystal structure of the N-terminal dimerization/oligomerization domain of TurB, an MvaT homologue in Pseudomonas putida KT2440. Our data identify two dimerization sites; the structure of the central dimerization site is almost the same as the corresponding region of H-NS, whereas the terminal dimerization sites are different. Our results reveal similarities and differences in dimerization and oligomerization mechanisms between H-NS and TurB.

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H-NS Oligomerization Domain Structure Reveals the Mechanism for High Order Self-association of the Intact Protein

Cited by 124 publications

References 29 publications

Modulation of Rho-Dependent Transcription Termination in Escherichia coli by the H-NS Family of Proteins

Modulation of Rho-Dependent Transcription Termination in Escherichia coli by the H-NS Family of Proteins

N9L and L9N mutations toggle Hha binding and hemolysin regulation by Escherichia coli and Vibrio cholerae H‐NS

Structural similarities and differences in H‐NS family proteins revealed by the N‐terminal structure of TurB in Pseudomonas putida KT2440

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