View details for Web of Science ID 000165066300004. Three Caulobacter crescentus heat-shock proteins were shown to be immunologically related to the Escherichia coli heat-shock proteins GroEL, Lon and DnaK. Cell division yields dissimilar daughter cells: a stalked cell and a swarmer cell that assembles several pili at the flagellated cell pole. 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. 2007: 506506. Stalked cells that developed directly from swarmer cells showed that same phospholipid profile as the swarmer cells. The genes that encode the components and regulatory proteins of the Caulobacter crescentus flagellum are transcribed at specific times in the cell cycle. The availability of CcrM is controlled in two ways: (1) the ccrM gene is transcribed only in the predivisional. The phage is similar to the Escherichia coli RNA phages in that it (i) sediments at an S(20, w) of 70.6S, (ii) is composed of a single molecule of single-stranded RNA and a protein coat, (iii) contains two structural proteins, and (iv) apparently contains the genetic capacity to code for a coat protein subunit, a maturation-like protein, and an RNA polymerase. By combining insights from multiple systems, its possible to identify the detailed molecular basis of many interesting evolutionary differences, including classic traits and diseases that affect millions of people around the world. View details for Web of Science ID A1976CE95700078. This result allowed us to deduce that the mechanism of fatty acid desaturation in C. crescentus is anaerobic, as it is in E. coli. On the other hand, several differences were found between the C. crescentus and E. coli RNA polymerases with respect to their interaction with Caulobacter phage phiCdl DNA. liqunyu2@illinois.edu A., Ryan, K. R., Shapiro, L., McAdams, H. H. The bifunctional FtsK protein mediates chromosome partitioning and cell division in Caulobacter, DnaA couples DNA replication and the expression of two cell cycle master regulators, Cytokinesis signals truncation of the PodJ polarity factor by a cell cycle-regulated protease. These basal bodies have five rings threaded on a rod. We have partially purified the ftr-binding proteins, and we show that they require the same enhancer sequences for binding as are required for transcriptional activation. These compounds were identified by screening for inhibitors against Caulobacter crescentus CcrM, an essential DNA methyltransferase from gram negative alpha-proteobacteria. View details for DOI 10.1111/j.1365-2958.2011.07698.x, View details for Web of Science ID 000292567200009, View details for PubMedCentralID PMC3137890. An unusual polar organelle that mediates directed motility on solid surfaces is found in the nonpathogenic bacterium Myxococcus xanthus. Stalked cells, which are actively engaged in DNA replication, have three or four SMC foci per cell. Two lines of evidence are presented which show that an RNase III activity functions as a processing enzyme in C. crescentus. View details for Web of Science ID 000259631900035, View details for PubMedCentralID PMC2566184. Asymmetric cell division in Caulobacter crescentus and sporulation in Bacillus subtilis are used as paradigms for the control of the cell cycle and cellular morphogenesis in bacterial cells. Even though NAPs affect DNA-related processes differently, all of them have to oligomerize and bind DNA for their function. Proteins are positioned at particular sites in bacteria, including the cell pole, the incipient division plane, and the septum. emw@med.unc.edu Letts, V., Shaw, P., Shapiro, L., Henry, S. INVITRO TRANSCRIPTION OF THE EARLY REGION OF CAULOBACTER PHAGE PHI-CD1 DEOXYRIBONUCLEIC-ACID BY HOST RNA-POLYMERASE, 3-DIMENSIONAL RECONSTRUCTION OF THE FLAGELLAR HOOK FROM CAULOBACTER-CRESCENTUS. Aminoacyl-transfer RNA (tRNA) synthetases, which catalyze the attachment of the correct amino acid to its corresponding tRNA during translation of the genetic code, are proven antimicrobial drug targets. Developmental biologist Lucy Shapiro, PhD, opened the second annual Discovery Innovation Awards event held on campus recently by sharing her personal research story. Nature Methods18, 945-952 (2021). View details for Web of Science ID A1990CL74300058, View details for Web of Science ID A1989AX26700001. What drives genomic innovation and diversity from bacterial to eukaryotes? Our partner, Stanford Blood Center, offers blood products to the Stanford community and beyond. Perhaps the periplasmic proteins are retained at the pole by the presence of the periseptal annulus (35). The parS/ParB chromosomal centromere is tethered to PopZ at one pole prior to the initiation of DNA replication. fliI encodes a 50-kDa polypeptide whose sequence is closely related to that of the Salmonella typhimurium FliI protein, an ATPase thought to energize the export of flagellar subunits across the cytoplasmic membrane through a type III protein secretion system. The flagellum and chemotaxis receptor are asymmetrically localized to a single pole in the predivisional cell by coordinated proteolysis and transcriptional regulation. 850 Harrison Ave 1st Floor Boston, MA 02118. View details for Web of Science ID 000165870600056. Frank Yang, lab member 2017-2019 PhD Candidate in Economics, Stanford Graduate School of Business, 2019-present BA Mathematics & Economics, Carleton College, 2017. Our results identify cell cycle changes in gene expression in response to carbon starvation that involve the prominent role of the FixK FNR/CAP family transcription factor and the CtrA cell cycle regulator. Graduate Student (joined @ 06/2017) Bioengineering. The formation of two distinct daughter cells upon division of the bacterium Caulobacter crescentus is the result of asymmetry in the predivisional cell, in part due to localization of both flagellar and chemotaxis proteins to the swarmer cell pole. Additionally, using ribosome profiling and LC-MS, we mapped translation start sites and coding regions with near complete coverage. Here, we report a strategy deployed by the asymmetrically dividing bacterium Caulobacter crescentus where a regulatory protein is programmed to perform distinct functions at the opposing cell poles. The Stanford Health Care (SHC) new 824,000 square-foot state-of-the-art hospital opened in 2019 with over 600 beds, making it one of the largest inpatient facilities in California. The pilus assembly protein TadZ (called CpaE in Caulobacter crescentus) is ubiquitous in tad loci, but is absent in other type IV pilus biogenesis systems. We propose that pole-specific control of PopZ function co-ordinates polar development and cell cycle progression by enabling independent assembly and tethering activities at the two cell poles. We report the identification of another C. crescentus heat shock operon containing two genes, hrcA (hrc for heat shock regulation at CIRCE elements) and a grpE homolog. A., Eckart, M. R., Shapiro, L. Three-Dimensional Super-Resolution Imaging of the RNA Degradation Machinery in Caulobacter Crescentus. These results contrast with RNase E and ribosome distribution in Escherichia coli, where RNase E colocalizes with the cytoplasmic membrane and ribosomes accumulate in polar nucleoid-free zones. B., Dingwall, A., Bryan, R., CHAMPER, R., Shapiro, L. POSITIONING OF GENE-PRODUCTS DURING CAULOBACTER CELL-DIFFERENTIATION, RATE, ORIGIN, AND BIDIRECTIONALITY OF CAULOBACTER CHROMOSOME-REPLICATION AS DETERMINED BY PULSED-FIELD GEL-ELECTROPHORESIS, ORGANIZATION AND TEMPORAL EXPRESSION OF A FLAGELLAR BASAL BODY GENE IN CAULOBACTER-CRESCENTUS, CONTROL OF SYNTHESIS AND POSITIONING OF A CAULOBACTER-CRESCENTUS FLAGELLAR PROTEIN. (5) Together, these regulatory proteins create a genetic circuit in which the cellular concentrations of CtrA and GcrA oscillate spatially and temporally to control daughter cell differentiation and cell cycle progression. This sequence of proteolytic events contributes to the asymmetric localization of PodJ isoforms to the appropriate cell pole. Space, Telecommunications and Radioscience Laboratory. In most cases of cellular asymmetry, bacteria are able to discriminate between the new pole and the old pole and to utilize this information for localization specificity. Observing crystallization using a time course of electron cryo-microscopy (Cryo-EM) imaging reveals a crystalline intermediate wherein N-terminal nucleation domains exhibit motional dynamics with respect to rigid lattice-forming crystallization domains. At the nonpermissive temperature, the cell cycle blocks prior to the de novo synthesis of flagella and chemotaxis proteins that normally occurs in the predivisional cell. Postdoctoral Fellow, Stanford University School of Medicine. Polar localization of the cytoplasmic CheA and CheW proteins is dependent on membrane-bound chemoreceptor. Although transcription from the smc promoter is induced during early S phase, a cell cycle transcription pattern previously observed with multiple DNA replication and repair genes, the SMC protein is present throughout the entire cell cycle. Caulobacter crescentus divides asymmetrically generating two distinct cell types at each cell division: a stalked cell competent for DNA replication, and a swarmer cell that is unable to initiate DNA replication until it differentiates into a stalked cell later in the cell cycle. Cell type-specific phosphorylation and proteolysis of a transcriptional regulator controls the G1-to-S transition in a bacterial cell cycle, Translation of the leaderless Caulobacter dnaX mRNA, Protein localization and cell fate in bacteria, Transcription of genes encoding DNA replication proteins is coincident with cell cycle control of DNA replication in Caulobacter crescentus. These results demonstrate that the chemotaxis methylation machinery is positionally biased toward one portion of the predivisional cell, and that the time of expression of a set of fla and che genes is correlated with the positioning of their gene products within the cell. Stem Cell Research. Chromosome segregation in Caulobacter cannot occur unless a dedicated parS guiding mechanism initiates movement. We demonstrated that the expression of a gene, flaD, that is involved in the formation of the flagellar basal body is under temporal control and is transcribed relatively early in the cell cycle, before the hook and flagellin genes are transcribed. The Caulobacter crescentus fliQ and fliR genes encode membrane proteins that have a role in an early step of flagellar biogenesis and belong to a family of proteins implicated in the export of virulence factors. We propose that during segregation PopZ sequesters free ParA and induces target-proximal regeneration of ParA DNA binding activity to enforce processive and pole-directed centromere segregation, preventing segregation reversals. Flagellated and non-flagellated vesicles were prepared from these cells by immunoaffinity chromatography and the level of MCPs that had been labeled either in vivo or in vitro with methyl-3H was determined. View details for Web of Science ID A1983RA96700072. Gahlmann, A., Ptacin, J. L., Grover, G., Quirin, S., von Diezmann, A. R., Lee, M. K., Backlund, M. P., Shapiro, L., Piestun, R., Moerner, W. E. Caulobacter chromosome in vivo configuration matches model predictions for a supercoiled polymer in a cell-like confinement. These sites overlap an essential DnaA box and a promoter in the origin that is essential for replication initiation. 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. We propose that a diverse repertoire of condensates can serve as control knobs to tune enzyme sequestration and reactivity in response to the metabolic state of bacterial cells. CtrA approximately P then accumulates and activates the transcription of cpdR, completing the regulatory loop, establishing an integrated network that controls a robust cell-cycle transition. Importantly, a small set of conserved ChpT residues promotes signaling crosstalk and contributes to the branched signaling that activates the master regulator CtrA while inactivating the CtrA degradation signal, CpdR. Accordingly, we identified, cloned, and sequenced a chromosomal locus, xylX, from Caulobacter crescentus which is required for growth on xylose as the sole carbon source and showed that transcription from a single site is dependent on the presence of xylose in the growth medium. View details for DOI 10.1016/j.tcb.2007.03.005, View details for Web of Science ID 000246939100005. Global transcription analysis of synchronized Caulobacter crescentus cells was used to identify 553 genes (19% of the genome) whose messenger RNA levels varied as a function of the cell cycle. CcrM is synthesized de novo late in the cell cycle, coincident with full methylation of the chromosome, and is then subjected to proteolysis prior to cell division. Here we report a novel assay to visualize pili by light microscopy that led to the purification of CAULOBACTER: pili and the isolation of a cluster of seven genes, including the major pilin subunit gene pilA. CtrA approximately P then blocks the reinitiation of replication while regulating the transcription of a large number of cell cycle-controlled genes. Progression of the Caulobacter cell cycle requires temporal and spatial control of gene expression, culminating in an asymmetric cell division yielding distinct daughter cells. Ranked in the top 10 for Neurology and Neurosurgical Care by US News and World Report, SHC is at the cutting edge of the latest treatments for neurological diseases. Moreover, initiation of DNA replication is allowed only once per cell cycle. Shapiro is an editor of Legal Theory and the Stanford Encyclopedia of Philosophy. Our laboratory is using genetic mapping, comparative sequence analysis, and functional tests to identify the genomic basis of classic evolutionary traits in vertebrates. View details for Web of Science ID 000228496100006. Although Drs. B.S. View details for Web of Science ID 000251336000004, View details for PubMedCentralID PMC2175322. The bacterial chemotaxis signal transducer MCP is an integral membrane receptor protein. Homology modelling of the N-terminal atypical receiver domain of CpaE indicates that it has a conserved protein-protein binding surface similar to that of the polar localization module of the social mobility protein FrzS, suggesting a similar function. Perturbing either MreB (with A22) or MreC (with depletion) causes GFP-Pbp2 to mislocalize to the division plane, indicating that each is necessary but not sufficient to generate a helical Pbp2 pattern. Laub, M. T., McAdams, T. H., Feldblyum, T., Fraser, C. M., Shapiro, L. Proteins on the move: dynamic protein localization in prokaryotes, tmRNAs that encode proteolysis-inducing tags are found in all known bacterial genomes: A two-piece tmRNA functions in Caulobacter. To study the relationship between phospholipid synthesis and organelle biogenesis in the dimorphic bacterium Caulobacter crescentus, auxotrophs have been isolated which require exogenous glycerol or glycerol 3-phosphate for growth when glucose is used as the carbon source. Intact bacterial flagella possessing a membrane-free hook and basal complex were purified from Caulobacter crescentus CB15, as well as from mutants which synthesize incomplete flagella. View details for Web of Science ID 000227028900009. Dahlberg, P. D., Sartor, A. M., Wang, J., Saurabh, S., Shapiro, L., Moerner, W. E. Integration of cell cycle signals by multi-PAS domain kinases. 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. Deoxyribonucleic acid-dependent ribonucleic acid (RNA) polymerase (EC 2.7.7.6) was purified from the dimorphic bacterium Caulobacter crescentus at three stages in development. The second region is adjacent to the hook and is approximately 10 nm in length. The deoxyribonucleic acid of the dimorphic bacterium Caulobacter crescentus contains a component that renatures with rapid, unimolecular kinetics. The examples of polar localization given here are from a variety of bacterial species and concern a disparate array of cellular functions. The CtrA/GcrA regulatory circuit controls expression of polar differentiation factors and the timing of DNA replication. Lucy Shapiro is a Professor in the Department of Developmental Biology at Stanford University School of Medicine where she holds the Virginia and D. K. Ludwig Chair in Cancer Research and is the Director of the Beckman Center for Molecular and Genetic Medicine. Despite their small size and lack of obvious intracellular structures, bacteria have a complex and dynamic intracellular organization. Isolation and characterization of a xylose-dependent promoter from Caulobacter crescentus. Childers, W., Xu, Q., Mathews, I. I., Mann, T. H., Blair, J. Our work demonstrates how nanoscale protein assemblies can modulate signal propagation with fine spatial resolution, and that in Caulobacter, this modulation serves to reinforce asymmetry and differential cell fate of the two daughter cells.
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