article|readcube | press, Researchers Make it Possible for Ultrasound to Reveal Gene Expression in the Body, Vilcek Foundation Prize Awarded to Mikhail Shapiro, CCE Postdoc Receives NIH Pathway to Independence Award, Mikhail Shapiro Wins Roger Tsien Award for Excellence in Chemical Biology, Program Brings Area High School Students, Teachers into Caltech Labs, Switching Brain Circuits On and Off Without Surgery, Mikhail Shapiro Selected as Camille Dreyfus Teacher-Scholar, Taking MRI Technology down to Micrometer Scales, Scientists Design Bacteria to Reflect Sonar Signals for Ultrasound Imaging, Biologists Give Bacteria Thermostat Controls, Designing Ultrasound Tools with Lego-Like Proteins, Newly Named Pew Scholar to Image Gut Bacteria with Sound Waves, Partnership with Heritage Medical Research Institute Will Augment Translational Medicine Research, Abedi Receives Fellowship for New Americans, Caltech Researchers Receive NIH BRAIN Funding, New Method Could Improve Ultrasound Imaging, x@caltech.edu; x=mikhail Immunoprecipitation of a DnaA'-beta-lactamase fusion protein showed that although expression occurs throughout the cell cycle, there is a doubling in the rate of expression just prior to the initiation of replication. Additionally, we summarize methods to use this fluoromodule for single protein imaging and super-resolution microscopy using stimulated emission depletion. Rate enhancement of protein crystallization by a discrete nucleation domain may enable engineering of kinetically controllable self-assembling 2D macromolecular nanomaterials. FtsZ is an essential bacterial GTPase that polymerizes at midcell, recruits the division machinery, and may generate constrictive forces necessary for cytokinesis. Topologically-guided continuous protein crystallization controls bacterial surface layer self-assembly. Macnab, J. Bacteriol. Transcript A appeared to be the major transcript since (a) it was the size of the entire 20% of the genome shown in vivo to code for the early phage mRNA, (b) it was one of the first transcripts synthesized at low enzyme-to-DNA molar ratios, and (c) it was synthesized in approximately 3 times the molar equivalent observed for the other transcripts. The ssrA deletion phenotype is not due to accumulation of stalled ribosomes, because the deletion is not complemented by a mutated version of SsrA that releases ribosomes but does not target proteins for degradation. MreB forms dynamic spirals in MreC-depleted cells, and MreC localizes helically in the presence of the MreB-inhibitor A22, indicating that each protein can form a spiral independently of the other. Spontaneous mutants have been isolated which are able to grow on galactose in the absence of exogenous cyclic nucleotides. Boehner, K., and J.T. RNA polymerase-binding studies with restriction fragments of the rRNA gene cluster revealed three regions which bound enzyme, and these regions were shown to contain transcription initiation sites. Twenty-three percent of the cell cycle-regulated promoters were found to be under the combinatorial control of two or more of the global regulators. We confirm that upstream sequences between -81 and -120 base-pairs are necessary for temporal activation, and that transcription also requires sequences at -26 to -46 base-pairs. CtrA plays key roles in asymmetric cell division and in the timing of chromosome replication. The CckA histidine kinase is known to contribute to CtrA phosphorylation. CcrM homologs are widespread in the alpha subdivision of gram-negative bacteria. However, protein crystallization as an evolutionary driver rationalizes S-layer diversity and raises the potential for biologically inspired self-assembling macromolecular nanomaterials. Analysis of the requirements for CtrA polar localization and CtrA proteolysis revealed that both processes require a motif within amino acids 1-56 of the CtrA receiver domain, and neither process requires CtrA phosphorylation. We have identified a periplasmic protease (PerP) that initiates the proteolytic sequence by truncating PodJ(L) to a form with altered activity (PodJ(S)). A high proportion of morphologically aberrant cells, and cells that have undergone an additional chromosome replication initiation, are found in this population. View details for DOI 10.1128/JB.185.16.4997-5002.2003, View details for Web of Science ID 000184692800037, View details for PubMedCentralID PMC166474. We show that the peptidoglycan transpeptidase Pbp2 also forms a helical pattern that partially colocalizes with MreC but not MreB. Genomically mined acoustic reporter genes for real-time in vivo monitoring of tumors and tumor-homing bacteria. collections of links for undergrads getting started in economics // work in progress Bio. The cellular localization of MipZ thus serves the dual function of positioning the FtsZ ring and delaying formation of the cell division apparatus until chromosome segregation has initiated. We describe the identification of 27 novel Caulobacter crescentus sRNAs by analysis of RNA expression levels assayed using a tiled Caulobacter microarray and a protocol optimized for detection of sRNAs. Chromosomal loci and many protein complexes are positioned at particular subcellular sites. 1973-1974 Stanford University Chengjian Mao Senior Researcher cmao@illinois.edu The first 258 amino acids of the N terminus are necessary and sufficient for targeting the protein to the division plane. All of the mutants in this cluster exhibited pleiotropic effects on the expression of other flagellar and chemotaxis functions, including the level of synthesis of flagellins, the hook protein and hook protein precursor, and the level of chemotaxis methylation. Although interactions between the chromosome and the cytoplasmic membrane are believed to be a functional component of the temporal regulation of DNA replication, the ability of this secA mutant to initiate replication at the nonpermissive temperature suggests that SecA-dependent events are not involved in this process. Although both enzymes protected the same sites on phiCdl DNA from cleavage with HincII, the E. coli enzyme was unable to form stable complexes with some phiCdl restriction fragments that formed stable complexes with the C. crescentus RNA polymerase. CtrA is a member of the response regulator family of two component signal transduction systems and is activated by phosphorylation. This origin also possesses three additional motifs that are unique to the C. crescentus origin of replication: seven 8-mer (GGCCTTCC) motifs, nine 8-mer (AAGCCCGG) motifs, and five 9-mer (GTTAA-n7-TTAA) motifs are present. Enhanced phase separation promotes the sequestration and activity of a client kinase enabling robust signaling and maintenance of viability under the stress posed by nutrient scarcity. View details for DOI 10.1128/JB.185.2.573-580.2003, View details for Web of Science ID 000180272600023, View details for PubMedCentralID PMC145339. The original point mutation is predicted to disrupt the stem structure in the 4.5 S RNA thus providing a rationale for the genetic basis of the LS439 phenotype. Professor of Biochemistry & Basic Medical Sciences, College of Medicine
Thus, two transcriptional feedback loops coupled to cell cycle-regulated proteolysis and phosphorylation of the CtrA protein result in the pattern of CtrA activity required for the temporal and spatial control of multiple cell-cycle events. Among the validated sRNAs, two are candidate transposase gene antisense RNAs. Translational efficiency (TE) was used as a metric for the relative rate of protein production from each mRNA. However, steady-state L-ring protein levels were dramatically reduced compared with those of wild type. Recently, single-molecule imaging and photocontrol have enabled superresolution optical microscopy of cellular structures beyond Abbe's diffraction limit, extending the frontier of noninvasive imaging of structures within living cells. Our results reveal a molecular mechanism that allows disparate environmental challenges to converge on a common pathway that results in a dormant state. Epistatic interactions between the genes accessed by the promoter probe and other flagellar loci were studied in double fla mutants generated by transducing the promoter-probe mutations into spontaneously derived second-site fla-mutant backgrounds. Time-lapse microscopy of the location of the chromosomal origin and 10 selected loci in the origin-proximal half of the chromosome showed that during DNA replication, as the replisome sequentially copies each locus, the newly replicated DNA segments are moved in chronological order to their final subcellular destination in the nascent half of the predivisional cell. To study cell-type-specific DNA initiation, chromosome replication was directly analyzed by pulsed-field gel electrophoresis. Single-molecule imaging in Caulobacter crescentus. 2014;2014: 4354-4357, Journal of wrist surgery -Van Nortwick, S., Berger, A., Cheng, R., Lee, J., Ladd, A. L.2013;2 (3): 263-270, Instructional course lectures -Ladd, A. L., Weiss, A. C., Crisco, J. J., Hagert, E., Wolf, J. M., Glickel, S. Z., Yao, J. In this paper, we report alterations in both the stalk and the flagellar structure that result from a mutation in the flagellar gene flbT. This binding activity was missing from strains containing mutations in flaO and flaW, two genes near the top of the flagellar hierarchy known to be required for hook operon transcription. These genes are controlled in a positive trans-acting hierarchy that reflects the order of assembly of the flagellum. However, many of the mechanistic details underlying these functions are unknown. B.S. Stanford Center on China's Economy and Institutions (SCCEI) Stanford Environmental and Energy Policy Analysis Center (SEEPAC) Stanford Digital Economy Lab; Stanford King Center on Global Development; Programs. These results demonstrate that MCP methylation is confined to that portion of the cell cycle when flagella are present. MAY 2011: Ed Green is named Searle Scholar for 2011 Ed was selected by the Kinship Foundation and the Chicago Community Trust as one of 15 Searle Scholars for 2011. Moreover moving the native ctrA gene to a position near the chromosomal terminus, which delays the conversion of the ctrA promoter from the fully to the hemimethylated state until late in the cell cycle, inhibited ctrA P1 transcription, and altered the time of accumulation of the CtrA protein and the size distribution of swarmer cells. The E. coli GroEL protein and the related protein from C. crescentus were also induced by treatment with hydrogen peroxide. In bacteria, studies of the cell cycle have focused largely on unsynchronized cells making it difficult to order the temporal events required for cell cycle progression, genome replication, and division. Caulobacter crescentus is widely used as a powerful model system for the study of prokaryotic cell biology and development. Full expression of fliX was found to be dependent on ctrA, and DNase I footprinting analysis demonstrated a direct interaction between CtrA and the fliX promoter. Two enzymes of galactose catabolism, galactose dehydrogenase and 2-keto-3-deoxy-6-phosphogalactonate aldolase, were shown to be inducible and independently regulated. von Diezmann, A., Lasker, K., Mann, T. H., Ahrens, D. G., Shapiro, L., Moerner, W. E. Probing Asymmetric Behavior of a Cell Cycle Regupatory Protein in Live Caulobacter using Single-Molecule Imaging, A Red Fluorescent Protein for Cryogenic Single-Molecule Superresolution Imaging. Strains bearing mutations in the pleA gene are pililess and non-flagellated. Stanford Report, February 1, 2013 President Obama presents the National Medal of Science to Stanford's Lucy Shapiro and Sidney Drell Bulman, George, and Robert Fairlie. Because of its critical role, CtrA activity is temporally regulated by multiple mechanisms including phosphorylation and ClpXP-dependent degradation of CtrA. The CtrA master transcriptional regulator is a central control element in Caulobacter cell cycle progression and polar morphogenesis. Double-stranded side branches between 100 and 600 nucleotide pairs in length were visible in electron micrographs of rapidly reassociating deoxyribonucleic acid isolated by hydroxyapatite chromatography. This directional movement of labeled MreB in the growing polymer provides an indication that, like actin, MreB monomers treadmill through MreB filaments by preferential polymerization at one filament end and depolymerization at the other filament end. Dynamic protein localization, phosphorelay signaling cascades, and spatially and temporally controlled proteolysis are overlayed on the transcription network that controls cell cycle progression and cell differentiation. A highly ordered chromosome structure, established while DNA replication and partitioning are in progress, is maintained and propagated during growth. Hurt RC#, Buss MT#, Duan M#, Wong K, You MY, Sawyer DP, Swift MB, Dutka P, Barturen-Larrea P, Mittelstein DR, Jin Z, Abedi MH, Farhadi A, Dephande R, Shapiro MG*. rRNA genes of Caulobacter crescentus CB13 were isolated and shown to be present in two gene clusters in the genome. PMID 15528588. We show that DnaA coordinates DNA replication initiation with cell cycle progression by acting as a global transcription factor. Collectively, our findings reveal an unsuspected level of environmental regulation of cell wall protein behavior that is likely linked to an ecological adaptation. Caulobacter crescentus undergoes an asymmetric cell division controlled by a genetic circuit that cycles in space and time. Our goal is to identify and understand the pathways that govern organogenesis of the pancreas, a vital organ with endocrine and exocrine functions. Interested applicants should visit https://facultypositions.stanford.edu/en-us/job/493432 for our full ad and more information about how to apply. While PodJS has a specific temporal and spatial address, MmpA is present throughout the cell cycle; furthermore, periplasmic fusion to mRFP1 suggested that MmpA is uniformly distributed around the cell. To understand the differential regulation of CtrA-controlled genes, we compared the expression of two of these genes, the fliQ flagellar gene and the ccrM DNA methyltransferase gene. Spatial organization and dynamics of RNase E and ribosomes in Caulobacter crescentus. The CtrA protein, a member of the response regulator family of the two-component signal transduction system, controls multiple cell cycle processes in Caulobacter and is present in swarmer cells but absent from stalked cells. 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