Fis binding to the recombinational enhancer of the Hin DNA inversion system.

Bruist, Michael F.; Glasgow, Anna C.; Johnson, Reid C.

doi:10.1101/gad.1.8.762

Cited by 98 publications

(122 citation statements)

References 35 publications

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“…These suggest that Fis also binds quantitatively and qualitatively well to the enhancer regardless of superhelical handedness. These data also suggest that the binding of Fis to enhancer is not as stable as shown in gel mobility shift assays (34). The reason for this is, we believe, that the MfeI reaction buffer in which the Fis-binding assay was performed was radically different from that used in gel mobility shift assays.…”

Section: Fis Binds To the Enhancer On (ϩ)Pkh336 -One Of The Fisbindinmentioning

confidence: 60%

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Hin-mediated Inversion on Positively Supercoiled DNA

Lim¹,

Lee²,

Jaxel³

et al. 1997

Journal of Biological Chemistry

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Hin recombinase requires negatively supercoiled DNA for an efficient inversion. We have generated positively supercoiled plasmid DNA using reverse gyrase from Sulfolobus shibatae and subjected it to the Hinmediated inversion reaction. Both Hin and Fis showed the same DNA binding activity regardless of the superhelical handedness of the substrate plasmid. However, inversion activity on positively supercoiled DNA was less than 1% of negatively supercoiled DNA. Assays designed to probe steps in inversion, showed that on positively supercoiled DNA, Hin was able to cleave the recombination sites with the same efficiency shown on negatively supercoiled DNA but was not able to exchange the cleaved DNA. Based on the theoretical differences between positive and negative supercoiling, our data may suggest that unwinding of the double helix at recombination sites is needed after DNA cleavage for strand exchange to occur.

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Section: Fis Binds To the Enhancer On (ϩ)Pkh336 -One Of The Fisbindinmentioning

confidence: 60%

“…* -3Ј sequence that contains the guanine residue (G * ) critical in Fis binding (34) and can be cleaved by MfeI restriction enzyme. pKH336 has an enhancer sequence between the hix sites.…”

Section: Fis Binds To the Enhancer On (ϩ)Pkh336 -One Of The Fisbindinmentioning

confidence: 99%

Hin-mediated Inversion on Positively Supercoiled DNA

Lim¹,

Lee²,

Jaxel³

et al. 1997

Journal of Biological Chemistry

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“…Two accessory proteins, Fis (factor for inversion simulation) and HU increase in vitro recombination rates over 100-fold relative to reactions performed with Hin alone (Johnson et al, 1986). Fis dimers bind two sites within the cis-acting recombinational enhancer (Johnson et al, 1986;Koch and Kahmann, 1986;Bruist et al, 1987a). The next step in the inversion reaction is the formation of a nucleoprotein complex known as the invertasome (Heichman and Johnson, 1990).…”

Section: Introductionmentioning

confidence: 99%

Role of arginine‐43 and arginine‐69 of the Hin recombinase catalytic domain in the binding of Hin to the hix DNA recombination sites

Adams

Nanassy

Johnson

et al. 1997

Molecular Microbiology

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SummaryThe Hin recombinase mediates the site-specific inversion of a segment of the Salmonella chromosome between two flanking 26 bp hix DNA recombination sites. Mutations in two amino acid residues, R43 and R69 of the catalytic domain of the Hin recombinase, were identified that can compensate for loss of binding resulting from elimination of certain major and minor groove contacts within the hix recombination sites. With one exception, the R43 and R69 mutants were also able to bind a hix sequence with an additional 4 bp added to the centre of the site, unlike wild-type Hin. Purified Hin mutants R43H and R69C had both partial cleavage and inversion activities in vitro while mutants R43L, R43C, R69S, and R69P had no detectable cleavage and inversion activities. These data support a model in which the catalytic domain plays a role in DNA-binding specificity, and suggest that the arginine residues at positions 43 and 69 function to position the Hin recombinase on the DNA for a step in the recombination reaction which occurs either at and/or prior to DNA cleavage.

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“…DNase I Footprint Assay-DNase I footprint assays were performed as previously described (14). End-labeled DNA probes were generated by PCR with one 5Ј-radiolabeled primer and a second nonradiolabeled primer.…”

Section: Methodsmentioning

confidence: 99%

The CTXϕ Repressor RstR Binds DNA Cooperatively to Form Tetrameric Repressor-Operator Complexes

Kimsey

Waldor

2004

Journal of Biological Chemistry

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CTX is a filamentous bacteriophage that encodes cholera toxin and integrates into the Vibrio cholerae genome to form stable lysogens. In CTX lysogens, gene expression originating from the rstA phage promoter is repressed by the phage-encoded repressor RstR. The N-terminal region of RstR contains a helix-turn-helix DNA-binding element similar to the helix-turn-helix of the cI/Cro family of phage repressors, whereas the short C-terminal region is unrelated to the oligomerization domain of cI repressor. Purified His-tagged RstR bound to three extended 50-bp operator sites in the rstA promoter region. Each of the RstR footprints exhibited a characteristic staggered pattern of DNase I-accessible regions that suggested RstR binds DNA as a dimer-ofdimers. In gel permeation chromatography and crosslinking experiments, RstR oligomerized to form dimers and tetramers. RstR was shown to be tetrameric when bound to operator DNA by performing mobility shift experiments with mixtures of RstR and a lengthened active variant of RstR. Binding of RstR to the high affinity O1 site could be fit to a cooperative model of operator binding in which two RstR dimers associate to form tetrameric RstR-operator complexes. The binding of RstR dimers to the left or right halves of O1 operator DNA was not observed in mobility shift assays. These observations support a model in which protein-protein contacts between neighboring RstR dimers contribute to strong operator binding.The molecular mechanisms that regulate bacteriophage lysogeny have been most extensively studied in phage lambda and its close relatives that infect Escherichia coli and Salmonella enterica serovar Typhimurium (1, 2). For these bacteriophages, the bistable switch from lysogenic to lytic development involves the interplay of two antagonistic transcriptional repressors, CI and Cro in the case of , that bind to the same set of regulatory sites in the bacteriophage control regions. A similar regulatory network controls lysogeny in Ø80 (3), HK022 (4), and the unrelated bacteriophage P2 (5). However, the regulation of lysogeny in a wide variety of other bacteriophages has not been investigated.CTX encodes the genes for cholera toxin, the virulence factor primarily responsible for the watery diarrhea characteristic of the disease cholera. Nontoxigenic strains of Vibrio cholerae can be readily transduced to toxin-producing strains by lysogenization with CTX, a process known as lysogenic conversion (6). Like other filamentous bacteriophage, CTX does not have a truly lytic growth phase. However, CTX lysogens exhibit several characteristics found in other temperate bacteriophages: 1) CTX lysogens contain the CTX prophage stably integrated into the V. cholerae genome and 2) the CTX prophage expresses a transcriptional repressor, RstR, that represses the expression of CTX replication genes and provides immunity to secondary infection by CTX (7,8).CTX shares genetic and morphological similarities with the E. coli filamentous bacteriophage fd. Similarities in gene sequence and gene orde...

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Fis binding to the recombinational enhancer of the Hin DNA inversion system.

Cited by 98 publications

References 35 publications

Hin-mediated Inversion on Positively Supercoiled DNA

Hin-mediated Inversion on Positively Supercoiled DNA

Role of arginine‐43 and arginine‐69 of the Hin recombinase catalytic domain in the binding of Hin to the hix DNA recombination sites

The CTXϕ Repressor RstR Binds DNA Cooperatively to Form Tetrameric Repressor-Operator Complexes

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