Purification and DNA-binding properties of the Cro-type regulatory repressor protein Cng encoded by the Lactobacillus plantarum phage φg1e

Kakikawa, Makiko; Ohkubo, Shingo; Sakate, Toshihide; Sayama, Michio; Taketo, Akira; Kodaira, Ken-Ichi

doi:10.1016/s0378-1119(00)00146-3

Cited by 14 publications

(4 citation statements)

References 15 publications

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“…The absence of detectable interaction between O3 and φ11 CI (as evident from both the gel shift and footprint assays) is quite unexpected because the primary immunity regions of phages λ [1], P22 [12], 434 [8], A2 [27], φg1e [28], HK022 [6] and N15 [29] bear more than two CI binding sites. Lactococcal phage Tuc2009 [30] and S. aureus φSa3ms [17], however, bear two CI binding sites, similar to that of φ11 in the cI‐cro intergenic region.…”

Section: Resultsmentioning

confidence: 99%

Physicochemical properties and distinct DNA binding capacity of the repressor of temperate Staphylococcus aureus phage φ11

Ganguly¹,

Das²,

Bandhu

et al. 2009

The FEBS Journal

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The basic regulatory elements that most temperate phages use for the establishment and maintenance of their lysogeny are the phage-encoded repressor and the cognate operator DNA [1][2][3][4][5][6][7][8][9][10][11][12]. A temperate phage generally enters into the lysogenic life cycle once its repressor inhibits the transcription of the phagespecific lytic genes from the early promoter by binding to the overlapped operator DNA. Repressors of the temperate phages, although varying greatly in size and in primary sequence level, mostly harbor a DNA binding domain and an oligomerization domain. The size and type of the operator DNAs also vary from phage to phage. Although some repressors bind to operators with dyad symmetry [1,[5][6][7][8][9] or operators with direct repeats [10], other repressors bind to asymmetric operators [2,3,[11][12][13] to establish lysogeny. Interestingly, the repressor of Vibrio cholerae phage CTX/ binds to extended operators, stopping lytic growth, as well as ensuring lysogeny of this phage [4]. Although these regulatory elements have enriched both basic and applied molecular biology enormously, they have not been cloned from most temperate phages or characterized in any depth.The temperate Staphylococcus aureus phage /11 [14] harbors the cI and cro genes in a divergent orientation to that in lambdoid phages [1,8]. The sequence of the immunity region of /11, however, differs significantly from those of the lambdoid phages and other temper- The repressor protein and cognate operator DNA of any temperate Staphylococcus aureus phage have not been investigated in depth, despite having the potential to enrich the molecular biology of the staphylococcal system. In the present study, using the extremely pure repressor of temperate Staphylococcus aureus phage /11 (CI), we demonstrate that CI is composed of a-helix and b-sheet to a substantial extent at room temperature, possesses two domains, unfolds at temperatures above 39°C and binds to two sites in the /11 cI-cro intergenic region with variable affinity. The above CI binding sites harbor two homologous 15 bp inverted repeats (O1 and O2), which are spaced 18 bp apart. Several guanine bases located in and around O1 and O2 demonstrate interaction with CI, indicating that these 15 bp sites are used as operators for repressor binding. CI interacted with O1 and O2 in a cooperative manner and was found to bind to operator DNA as a homodimer. Interestingly, CI did not show appreciable binding to another homologous 15 bp site (O3) that was located in the same primary immunity region as O1 and O2. Taken together, these results suggest that /11 CI and the /11 CI-operator complex resemble significantly those of the lambdoid phages at the structural level. The mode of action of /11 CI, however, may be distinct from that of the repressor proteins of k and related phages.Abbreviations CI, repressor of temperate Staphylococcus aureus phage /11; CTD, C-terminal domain; DMS, dimethyl sulfate; DTNB, 5,5¢-dithiobis-(2-nitrobenzoic acid); NTD, N-terminal domain.

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Section: Resultsmentioning

confidence: 99%

Physicochemical properties and distinct DNA binding capacity of the repressor of temperate Staphylococcus aureus phage φ11

Ganguly¹,

Das²,

Bandhu

et al. 2009

The FEBS Journal

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“…Among other orfs of particular interest, the cpg gene is expressed through the promoter P L and encodes a 132-aa protein similar to SOSrelated repressors, while cng, under the control of the promoter P R oriented in the opposite direction, encodes a Cro-like repressor (88 aa). Both proteins can bind to operator-like GATAC boxes but in slightly different way (101,102). As shown by footprint assays, Cng does not completely cover two of the seven boxes identified in the phig1e genome (namely Gb4 and Gb6).…”

Section: Lactobacillus Plantarum Phages 441 Phig1ementioning

confidence: 94%

Bacteriophages of Lactobacillus

Villion

Moineau²

2009

Front Biosci

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In this review, we are listing Lactobacillus phages that have been reported in peer-reviewed articles published since 1960. Putative phages that are defective or have not been shown to be infectious, such as phage-like particles, are not discussed. Our literature searches led to the identification of 231 Lactobacillus phages, 186 of which have been observed by electron microscopy, with 109 belonging to the Siphoviridae family, 76 to the Myoviridae family, and 1 to the Podoviridae family. Model phages infecting Lb delbrueckii, casei, rhamnosus, plantarum, and gasseri are highlighted, as well as prophages of Lactobacillus hosts. To date, nine complete Lactobacillus phage genomes are available for comparisons and evolution studies. Features such as phage receptors and endolysins are also reviewed, as well as phage-derived genetic tools. Lactobacillus phage research has progressed significantly over the past decade but a thorough understanding of their biology is still lacking. Because of the risks they represent and the knowledge gaps that need to be filled, the outlook for research on Lactobacillus phages is bright.

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“…The molecular basis for the lysislysogeny switch is well understood for , but the exact mechanisms for sensing environmental conditions and how the signaling routes are coupled to the molecular regulation mechanisms are not well defined. Recently, regulatory mechanisms involved in the lysis-lysogeny decision have been reported for several lactic acid bacterial phages (3,4,6,15,18,20,23,24,30,35,45), and similarities exist between the regulatory mechanisms of lysogenic lactic acid bacterial phages and .…”

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confidence: 99%

Effects of Diverse Environmental Conditions on φLC3 Prophage Stability in Lactococcus lactis

Lunde¹,

Aastveit²,

Blatny³

et al. 2005

Appl Environ Microbiol

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The effects of various growth conditions on spontaneous LC3 prophage induction in Lactococcus lactis subsp. cremoris IMN-C1814 was analyzed with a half fraction of a 4 4 factorial experimental design. The four factors included in the study were nutrient availability, acidity, osmolarity, and temperature, each applied at four levels. These environmental factors are related to the fermentation processes in the dairy industry, in which bacteriophage attacks on sensitive starter strains are a constant threat to successful fermentation processes. The frequency of spontaneous LC3 induction was determined by quantitative analyses of restored DNA attachment sites (attB) on the bacterial chromosomes in a population of lysogenic cells. Statistical analysis revealed that all four environmental factors tested affected LC3 prophage stability and that the environmental factors were involved in interactions (interactions exist when the effect of one factor depends on the level of another factor). The spontaneous LC3 induction frequency varied from 0.08 to 1.76%. In general, the induction frequency remained at the same rate or decreased when level 1 to 3 of the four environmental factors was applied. At level 4, which generally gave the least favorable growth conditions, the induction frequency was either unchanged, decreased, or increased, depending on the type of stress. It appeared that the spontaneous induction frequency was independent of the growth behavior of the host. It was the environmental growth conditions that were the decisive factor in induction frequency.

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Purification and DNA-binding properties of the Cro-type regulatory repressor protein Cng encoded by the Lactobacillus plantarum phage φg1e

Cited by 14 publications

References 15 publications

Physicochemical properties and distinct DNA binding capacity of the repressor of temperate Staphylococcus aureus phage φ11

Physicochemical properties and distinct DNA binding capacity of the repressor of temperate Staphylococcus aureus phage φ11

Bacteriophages of Lactobacillus

Effects of Diverse Environmental Conditions on φLC3 Prophage Stability in Lactococcus lactis

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