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cvhA Gene of Streptomyces hygroscopicus 10-22 Encodes a Negative Regulator for Mycelia Development

Acta Biochimica et Biophysica Sinica 2006年第4期38卷 271-280页

作者： Heng-An WANG~(1,2#) Lei QIN~(2#) Ping LU~(1*) Zhi-Xuan PANG~2 Zi-Xin DENG~(3*) Guo-Ping ZHAO~(2*) 1 Biotechnology Institute,School of Agriculture and Biology,Shanghai Jiaotong University,Shanghai 201101,China 2 Laboratory of Molecular Microbiology,Institute of Plant Physiology and Ecology,Shanghai Institute for Biological Sciences,Chinese Academy of Sciences,Shanghai 200032,China 3 Bio-X Life Science Research Center,Shanghai Jiaotong University,Shanghai 200030,China. BiotechnologyInstitute SchoolofAgricultureandBiologyShanghaiJiaotongUniversityShanghai201101China LaboratoryofMolecularMicrobiology InstituteofPlantPhysiologyandEcologyShanghaiInstituteforBiologicalSciencesChinese

A five-gene cluster cvhABCDE was identified from Streptomyces hygroscopicus *** thefirst gene of this cluster,cvhA encoded a putative sensor histidine kinase with a predicted sensor domainconsisting of two trans-membrane segments at the N-terminus and a conserved HATPasecdomain at the *** C-terminus polypeptide of CvhA expressed in Escherichia coli was purified and shown to beautophosphorylated with [γ-32P]ATP in *** phosphoryl group was acid-labile and basic-stable,whichsupported histidine as the phosphorylation *** obvious difference of mycelia development was ob-served between the null mutant of cvhA generated by targeted gene replacement and the wild-type parentalstrain 10-22 grown on solid soya flour medium with 2%-8% glucose or sucrose,but the cvhA mutant couldform much more abundant aerial mycelia and spores than the wild-type strain on solid soya flour mediumsupplemented with 6%-8% mannitol,6%-8% sorbitol,4%-6% mannose,or 4%-6% *** pheno-type was complemented by the cloned wild-type cvhA gene,and no difference was observed for growthcurves of the cvhA mutant and the wild strain in liquid minimal medium with the tested sugars at a concen-tration of 4%,6% and 8%.We thus propose that CvhA is likely a sensor histidine kinase and negativelyregulates the morphological differentiation in a sugar-dependent manner in *** 10-22.

关键词： sensor histidine kinase aerial mycelia sugar metabolism

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Structure-Function Analysis of Arabidopsis thaliana histidine kinase AHK5 Bound to Its Cognate Phosphotransfer Protein AHP1

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Molecular Plant 2013年第3期6卷 959-970页

作者： Johannes Bauer Kerstin Reiss Manikandan Veerabagu Michael Heunemann Klaus Harter Thilo Stehle Interfaculty Institute of Biochemistry University of Tubingen D-72076 Tubingen Germany Center for Plant Molecular Biology University of TObingen D-72076 T0bingen Germany Department of Pediatrics Vanderbilt University School of Medicine Nashville TN 37232 USA

The multi-step phosphorelay （MSP） system defines a key signal transduction pathway in plants and many eukaryotes. In this system, external stimuli first lead to the activation of a histidine kinase, followed by transfer of a phosphoryl group from the receiver domain of the kinase （HKRD） to downstream, cytosolic phosphotransfer proteins （HPs）. In order to establish the determinants of specificity for this signaling relay system, we have solved the first crystal structure of a plant HKRD, AHK5RD, in complex with one of its cognate HPs, AHP1. AHP1 binds AHK5RD via a prominent hydrogen bond docking ridge and a hydrophobic patch. These features are conserved among all AHP proteins, but differ significantly from other structurally characterized prokaryotic and eukaryotic HPs. Surface plasmon resonance experiments show that AHK5RD binds to AHP1-3 with similar, micromolar affinity, consistent with the transient nature of this signaling complex. Our correlation of structural and functional data provide the first insight, at the atomic level as well as with quantitative affinity data, into the molecular recognition events governing the MSP in plants.

关键词： multi-step phosphorelay phosphotransfer protein plant signaling sensor histidine kinase two-component system.

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