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1、张荣娟差异基因代号基因推断功能同源性RJ2Conserved protein(氧化还原酶位于同一个操纵元中，还没有关于此基因的研究)98%RJ4secAProbable preprotein translocase SecA subunit functions in protein export; 74%RJ6CgtA和ProB1Putative GTP-binding protein和Probable glutamate 5-kinase98%RJ7prkAPrkA; Putative Ser protein kinase（Signal transductionmechanisms）76%R
2、J9gltAtype II citrate synthase:（Citrate synthase activity was essential for nodule maintenance ）70%RJ14thiCputative thiamine biosynthesis protein68%RJ5fixRJ12Transcriptional regulator, LysR family 与1021相似性低但与其他物种比对也是这个蛋白总结-6个差异基因JY6B和JY6（下调）Disruption of the cgtA gene was lethal, demonstrating that
3、this gene is essential for cell growth. secAE.coli and Bacillus 功能已经明确， hydrolyzes ATP and an essential component of the protein translocation ATPase ；SecA Protein的ATP-binding site已经确定；SecA 识别并结合 preprotein-SecB complex, 结合到 inner-membrane anionic phospholipids, 插入 the membrane bilayer 以后，再与the prep
4、rotein translocator, SecY/SecE结合，从而将蛋白运输过膜.putativetransportproteintransmembranehypotheticalproteincbaSecBthiC文献分布：Sinorhizobium meliloti (2)R.etli(2)E.coli and else(15)Sinorhizobium 突变体库中有关于这个突变体的信息；Rhizobium etli：在其中表达thiCOGE，将增加其固氮效率；Thiamin biosynthesis in prokaryotes：其中涉及12个基因（ thiFSGH, I, and
5、dxs， thiC， thiE， thiD, thiM, thiL, and pdxK）突变体库查询内容prkA(1)hypothetical protein SMc01266 Bacillus subtilis ：prkA gene encoding a novel serine protein kinasecbaSMc00367（假定蛋白）Protein:putative oxidoreductase protein putative oxidoreductase protein RJ2RJ2360bpSMc00367Conserved protein(氧化还原酶位于同一个操纵元中, 同源
6、性98%, 还没有关于此基因的研究)98%JY6B和JY6（下调）/ obgEobgE/proB150S ribosomal protein L21 hypothetical protein gamma-glutamyl kinase gamma-glutamyl phosphate reductase nicotinic acid mononucleotide adenylyltransferase obgE (15篇E.coli most) (cgtA ，yhbZ, obgE）coding for an essential GTP-binding protein，染色体复制、分裂等过程中起
7、调节作用，可能作用于核糖体合成的最后一步。proB1（ Escherichia coli）Glutamate-5-kinase： proline（脯氨酸） 由 glutamate（谷氨酸盐）合成需要三步： catalyzed by glutamate 5-kinase (G5K), glutamyl 5-phosphate reductase (G5PR) and pyrroline 5-carboxylate reductase；脯氨酸plays an important role as an osmoprotectant in microorganismsGlutamate-5-kinas
8、e 的active site residues已确定The EMBO Journal vol.8 no.3 pp.961 -966, 1989SecA protein hydrolyzes ATP and is an essential component of the protein translocation ATPase of Escherichia coliCharacterization of a Bacillus subtilis SecA mutant protein deficient in translocation ATPase and release from the m
9、embrane Histidine Residues Are Involved in Translocation-Coupled ATP Hydrolysis by the SecA Protein MolMicrobiol.1993Apr;8(1):31-42.Characterization of a Bacillus subtilis SecA mutant protein deficient in translocation ATPase and release from the membrane-It is concluded that the GKT motif in the am
10、ino-terminal domain of SecA is part of the catalytic ATP-binding site. This site may be involved in the ATP-driven protein recycling function of SecA which allows the release of SecA from its association with precursor proteins, and the phospholipid bilayer. secASecAprotein:autoregulatedATPasecataly
11、singpreproteininsertionandtranslocationacrosstheEscherichiacoliinnermembranRecentinsightintothebiochemicalmechanismsofproteintranslocationinEscherichiacoliindicatesthatSecAATPaseisrequiredbothfortheinitialbindingofpreproteinstotheinnermembraneaswellassubsequenttranslocationacrossthisstructure.SecAap
12、pearstopromotetheseeventsbydirectrecognitionofthepreproteinorpreprotein-SecBcomplex,bindingtoinner-membraneanionicphospholipids,insertionintothemembranebilayerandassociationwiththepreproteintranslocator,SecY/SecE.ATPbindingappearstocontroltheaffinityofSecAforthevariouscomponentsofthesystemandATPhydr
13、olysispromotescyclingbetweenitsdifferentbiochemicalstates.Asacomponentlikelytocatalysearate-determiningstepinproteinsecretion,SecAsynthesisisco-ordinatedwiththeactivityoftheproteinexportpathway.ThisformofnegativeregulationappearstorelyonSecAproteinbindingtoitsmRNAandrepressingtranslationifconditions
14、ofrapidproteinsecretionprevailwithinthecell.AprecisebiochemicalschemeforSecA-dependentcatalysisofproteinexportandthedetailsofsecAregulationappeartobecloseathand.TheevolutionaryconservationofSecAproteinamongeubacteriaaswellasthegeneralrequirementfortranslocationATPasesinotherproteinsecretionsystemsar
15、guesforamechanisticcommonalityofallprokaryoticproteinexportpathways.prkACloning and characterization of the Bacillus subtilis prkA gene encoding a novel serine protein kinase. Cloning, expression, purification and characterization of the stress kinase YeaG from Escherichia coli.Characterization of t
16、he Bacillus subtilis thiC OperonInvolved in Thiamine BiosynthesisExpression of Thiamin Biosynthetic Genes (thiCOGE) and Productionof Symbiotic Terminal Oxidase cbb3 in Rhizobium etli These data show a direct relationship between expression of thiC and production of the cbb3 terminaloxidase. This is
17、consistent with the proposition that a purine-related metabolite, 5-aminoimidazole-4-carbox-amide ribonucleotide, is a negative effector of the production of the symbiotic terminal oxidase cbb3 in R. etli.Thiamin biosynthesis in prokaryotes Twelve genes involved in thiamin biosynthesis inprokaryotes
18、 have been identified and overexpressed. Ofthese, six are required for the thiazole biosynthesis (thiF-SGH, I, and dxs), one is involved in the pyrimidinebiosynthesis (thiC), one is required for the linking of thethiazole and the pyrimidine (thiE), and four are kinases(thiD, thiM, thiL, and pdxK). A
19、 conserved RNA structure (thi box) is involved in regulation of thiamin biosynthetic gene expression in bacteria(R. etli strain CE3)thiCThiamine limitation determines the transition fromaerobic tofermentative-like metabolism in Rhizobium etli CE3ThiC Is an Fe-S Cluster Protein That Requires AdoMet T
20、o Generate the-Amino-5-hydroxymethyl-2-methylpyrimidine Moiety in Thiamin SynthesisReaction of AdoMet with ThiC generates a backbone free radicalRole of the cgtA gene function in DNA replication of extrachromosomal elements in Escherichia coli(2003) It seems that DNA synthesis per se is not affected
21、 by CgtA, and that this protein might control replication initiation indirectly, by regulation of function(s) or production of one or more replication factors. In fact, we found that level of the host-encoded replication protein DnaA is significantly decreased in the cgtA mutant. This indicates that
22、 CgtA is involved in the regulation of dnaA gene expression. Overexpression of the cgtA (yhbZ, obgE) gene, coding for an essential GTP-binding protein, impairs the regulation of chromosomal functions in Escherichia coli.(2002)DNA replication defect in the Escherichia coli cgtA(ts) mutant arising fro
23、m reduced DnaA levels(2006) Obg/CtgA, a Signaling Protein That Controls Replication, Translation, and Morphological Development?(2006) The recent finding that the ObgE GTPase acts as a ated with Obg: the effects on bacterial replication (Foti replication checkpoint protein in Escherichia coli has et
24、 al., 2005). A new E. coli obgE mutant was isolated important implications.Involvement of the cgtA gene function in stimulation of DNA repair in Escherichia coli and Vibrio harveyi(2003)The Escherichia coli GTPase CgtAE Is Involved in Late Steps of Large Ribosome Assembly(2006) Mutations in CgtAE ca
25、use both polysome and rRNA processing defects; small- and large-subunit precursor rRNAs accu-mulate in a cgtAE mutant.The ObgE/CgtA GTPase influences the stringent response toamino acid starvation in Escherichia coli(2009)Functional analysis of the GTPases EngA and YhbZencoded by Salmonella typhimur
26、ium(2007)AbstractThe S. typhimurium genome encodes proteins, designated EngA and YhbZ, which have a high sequence identity with the GTPases EngA/Der and ObgE/CgtAE of Escherichia coli. The wild-type activity of the E. coli proteins is essential for normal ribosome maturation and cell viability. In o
27、rder to characterize the potential involvement of the Salmonella typhimurium EngA and YhbZ proteins in ribosome biology, we used high stringency affinity chromatography experiments to identify strongly binding ribosomal partner proteins. A combination of biochemical and microcalorimetric analysis wa
28、s then used to characterize these protein:protein interactions and quantify nucleotide binding affinities. These experiments show that YhbZ specifically interacts with the pseudouridine synthase RluD (KD 2 mM and 1:1 stoichiometry), and we show for the first time that EngA can interact with the ribo
29、somal structural protein S7. Thermodynamic analysis shows both EngA and YhbZ bind GDP with a higher affinity than GTP (20-fold difference for EngA and 3.8-fold for YhbZ), and that the two nucleotide binding sites in EngA show a 5.3-fold difference in affinity for GDP. We report a fluorescence assay
30、for nucleotide binding to EngA and YhbZ, which is suitable for identifying inhibitors specific for this ligand-binding site, which would potentially inhibit their biological functions. The interactions of YhbZ with ribosome structural proteins that we identify may demonstrate a previously unreported
31、 additional function for this class of GTPase: that of ensuring delivery of rRNA modifying enzymes to the appropriate region of the ribosome.Chromosome segregation control by Escherichia coli ObgE GTPase(2007)We propose a model in which ObgE is required to license chromosome segregation and subsequent cell cycle events.Glutamate-5-kinase from Escherichia coli: gene cloning, overexpression, purification and crystallization of the recombinant enzyme and preliminary X-ray studies(2005)The substrate glutamate and the feed-back inhibitor proline bind at overlapping sites(2006)
