One of these shares a promoter motif with several genes expressed at the swarmer-to-stalked cell transition; while another appears to be controlled by the CtrA global transcriptional regulator.
Collaboration: In-vivo Drug Evaluation, University of Colorado and Health Sciences Center
GENERATION OF POLARITY DURING CAULOBACTER CELL-DIFFERENTIATION, PHOSPHORYLATION OF THE BETA'-SUBUNIT OF RNA-POLYMERASE AND OTHER HOST PROTEINS UPON PHI-CD1 INFECTION OF CAULOBACTER-CRESCENTUS, ORGANIZATION AND NUCLEOTIDE-SEQUENCE ANALYSIS OF AN RIBOSOMAL-RNA AND TRANSFER-RNA GENE-CLUSTER FROM CAULOBACTER-CRESCENTUS.
We show here that the ClpXP protease, responsible for the degradation of multiple bacterial proteins, is dynamically localized to specific cellular positions in Caulobacter where it degrades colocalized proteins. We analyzed the adaptive response of C. crescentus swarmer cells to carbon starvation and found that there was a block in both the swarmer-to-stalked cell polar differentiation program and the initiation of DNA replication.
The first region is the hook.
We demonstrate here that CcrM is an important target of the Lon protease pathway in C. crescentus.
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CtrA activity in the cell cycle is controlled both transcriptionally and by phosphorylation. Our laboratory is interested in the growth, development and integrity of animal tissues.
Its chromosome replication origin (Cori) may be prototypical of the large and diverse class of alpha-proteobacteria.
We have isolated the sigma 54 gene (rpoN) and describe here a novel role for this alternative sigma-factor in cell differentiation: It is required for the biogenesis of both polar structures, and the disruption of the rpoN gene results in aberrant cell division. In order to study the regulation of these genes, plasmids were constructed that contain either an intact flaYE region or deletions in the region of flaY.
SpoT, however, was not required for the carbon starvation block of the swarmer-to-stalked cell polar differentiation program.
The proteolytic substrate PodJ(L) is a polar factor that recruits proteins required for polar organelle biogenesis to the correct cell pole at a defined time in the cell cycle. Instead, separate proteins differing in enzymatic activity were detected, analogous to the beta-oxidation enzymes that have been isolated from Clostridium acetobutylicum and from mitochondria of higher eucaryotes. Mutations that inhibited dynamic PopZ localization inhibited the recruitment of other factors to cell poles.
A simple antibody-based bioassay using fluorescently tagged proteins demonstrates the encoding strategy in biologically relevant media. Purucker, M., Bryan, R., Amemiya, K., Ely, B., Shapiro, L. CHARACTERIZATION OF THE PROTEINS OF THE CAULOBACTER-CRESCENTUS FLAGELLAR FILAMENT - PEPTIDE ANALYSIS AND FILAMENT ORGANIZATION.
While asymmetric division in Caulobacter normally yields larger stalked and smaller swarmer daughters, we observe a loss of asymmetric size distribution among daughter cells when clpA is depleted from a strain in which FtsZ is constitutively produced.
The assembly of a functional flagellum in the bacterium Caulobacter crescentus requires the protein products of approximately 30 genes expressed in a temporally discrete and spatially distinct manner.
Further, we find that a mutation to glycine of two conserved aspartic acid residues that are important for nucleotide hydrolysis in other members of the actin superfamily abolishes robust midcell recruitment of MreB but supports a normal rate of growth. We seek to understand the mechanisms that regulate self-renewal, proliferation and differentiation in adult stem cell linages.
With one method, the M ring makes a snug contact with the S ring and is often capped by an axial button, a new component apparently distinct from the M ring.
Accurate replication and segregation of the bacterial genome are essential for cell cycle progression. The S ring has a triangular cross section, the sides of the triangle abutting the E ring, the rod and the M ring.
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.
Caulobacter crescentus is a premier model organism for studying the molecular basis of cellular asymmetry. Ultrasensitive ultrasound imaging of gene expression with signal unmixing.
Schrader, J. M., Li, G., Zhou, B., Weissman, J. S., Shapiro, L. Quantifying the Spatial Organization of Bacterial Ribosomes using Three-Dimensional Super-Resolution Microscopy.
Tn5 insertion mutants unable to form colonies when oleic acid was the sole carbon source were isolated.
SciP is cell cycle-controlled and co-conserved with the global cell cycle regulator CtrA in the -proteobacteria. Caulobacter goes to great lengths to control the time and place of the activity of this critical regulatory factor during the cell cycle. Postdoctoral Fellow, Stanford University School of Medicine, A.B. Kang, Kim and Shin used to work with Dr. Park, they became independent scientists with own research projects.
In addition, increasing the cellular content of sigma32 by introducing a plasmid-encoded copy of rpoH induced DnaK expression in C. crescentus cultures grown at 30 degrees C. The C. crescentus rpoH gene was transcribed from either of two heat shock consensus promoters.
Caulobacter crescentus cell division is asymmetric and yields distinct swarmer cell and stalked cell progeny.
The bacterium Caulobacter crescentus undergoes an asymmetric cell division resulting in the formation of two different daughter cells, a motile swarmer cell and a nonmotile stalked cell. We demonstrate that SciP binds to DNA at a motif distinct from the CtrA binding motif that is present in the promoters of genes co-regulated by SciP and CtrA.
The expression of these class II genes initiates assembly of the flagellum just prior to activation of the ccrM promoter in the predivisional cell.
The Tol-Pal complex is required to maintain the position of the transmembrane TipN polar marker, and indirectly the PleC histidine kinase, at the cell pole, but it is not required for the polar maintenance of other transmembrane and membrane-associated polar proteins tested. Such organization is transmitted from one generation to the next by progressive segregation of daughter chromosomes and anchoring of DNA to the cell envelope.
A protein kinase activity is induced early after infection of Caulobacter crescentus by the DNA phage phiCd1.
View details for Web of Science ID A1994MQ86600007. The experience inspired him to apply after he graduated in 2013, and he was accepted. Learn how we are healing patients through science & compassion, Stanford team stimulates neurons to induce particular perceptions in mice's minds, Students from far and near begin medical studies at Stanford. The cell cycle arrest therefore results from nutritional deprivation and is analogous to the general control system exhibited by yeast (Hartwell, Bacteriol.
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Many important biological processes occur deep inside living organisms. Using genetic screens and cellular approaches in zebrafish, we aim to discover new genes with essential functions in glial cells, define new animal models of important disorders in humans, and provide new avenues toward therapies for injury and disease of the nervous system.
Rather, the mutants appeared to have defects relating to the complex coordination of membrane biogenesis and cell cycle events in C. crescentus. Caulobacter requires micromolar concentrations of calcium for normal growth and development. ddurak2@illinois.edu
The promoter sequence does not resemble that recognized by any known bacterial sigma factor.
Bacterial chromosome origins of replication.
Biophysics of neuromodulation with ultrasound and other forms of energy.
Here, we report that in Caulobacter, a hipA2-encoded bacterial toxin contributes to bacterial persistence by manipulating intracellular amino acid balance. cmao@illinois.edu
Transcription activation of flbN in C. crescentus involves the combination of several elements: the NifA-like site is required for full activation, and other sequence elements 5' to the promoter and 3' to the transcription start site are necessary for the correct time of transcription initiation.
The start site of the fliLM operon lies 166 bp from the divergently transcribed flaCBD operon that encodes several basal body genes. Our aim is to identify and characterize systems that influence the interplay among genetic variation, phenotypic diversity, and environmental fluctuations at the molecular level, integrating our findings to gain insight into complex cellular systems. Amemiya, K., Bellofatto, V., Shapiro, L., Feingold, J. Join the team at Stanford Anatomic Pathology and Clinical Labs. This 37,000 Mr heat-shock protein might be related to the E. coli 32,000 Mr heat-shock sigma subunit.
SpoT is a bifunctional synthase/hydrolase that controls the steady-state level of the stress-signaling nucleotide (p)ppGpp, and carbon starvation caused a SpoT-dependent increase in (p)ppGpp concentration. In the bacterium Caulobacter crescentus, many cellular processes are temporally regulated with respect to the cell cycle, and the genes required for these processes are expressed immediately before the products are needed. This result establishes the editing site as a bona fide target for aminoacyl-tRNA synthetase inhibitors.
The cell-cycle control has a cyclical genetic circuit composed of four regulatory proteins with tight coupling to processive chromosome replication and cell division subsystems.
At the swarmer-to-stalked cell transition and in the stalked compartment of the predivisional cell, CtrA is localized to the cell pole just before its degradation.
czhang8@illinois.edu
Models for regulation of Caulobacter early flagellar promoters are discussed in which RNA polymerase containing a novel sigma subunit interacts with an activation factor bound to the central region of the promoter.
Antigen-antibody complex formation occurring within a vector-phage plaque can be used to detect the production of a specific protein from an amplified gene. The parS sites, a pair of short contiguous sequence elements known to be involved in chromosome segregation, are positioned at one pole, where they anchor the chromosome to the cell and contribute to the formation of a compact chromatin conformation. She is a professor of Developmental Biology at the Stanford University School of Medicine.
One-third of these target genes encode putative TonB-dependent receptors, suggesting CrfA plays a role in the surface modification of C. crescentus, facilitating the uptake of nutrients during periods of carbon starvation. Although the newly replicated origin regions of the chromosome are rapidly moved to opposite cell poles by an active process, the replisome appears to be an untethered replication factory that is passively displaced towards the center of the cell by the newly replicated DNA. In this report we identify a cluster of genes encoding basal body components and describe their transcriptional regulation.
These data suggest a more prevalent use of the Shine-Dalgarno sequence for ribosome pausing rather than translation initiation in C. crescentus.
Trisha went on to join Lucy Shapiro's lab for her thesis research.
Thus, fatty acid degradation by the beta-oxidation pathway is constitutive in C. crescentus and is only mildly affected by growth in the presence of glucose. View details for Web of Science ID A1983QR08100025. | NYT News.
We isolated 35 unique A22-resistant Caulobacter strains with single amino acid substitutions near the nucleotide binding site of MreB. A specialized protein secretion pathway is used by some Gram-negative bacterial pathogens for delivery of virulence factors directly into mammalian host cells. Among the validated sRNAs, two are candidate transposase gene antisense RNAs.
The availability of temperature-sensitive mutants blocked at various stages of development permits access to both questions. Thus, we propose that the Caulobacter chromosomal origins have specific cellular addresses and that the SMC protein plays important roles in maintaining chromosome structure and in partitioning.
Interestingly, M. xanthus, which has nozzles at both poles, can reverse direction by closing one nozzle and opening the other in response to end-to-end interactions between cells.
Independent mutations in the conserved sequence that lies between the -10 and -35 regions increased transcription, suggesting that a repressor may bind at this site.
Thanks to all the lab members, collaborators and friends who joined us for the annual Shapiro Lab beach party in Oceanside, CA!
Chen, S. L., Lee, W., Hottes, A. K., Shapiro, L., McAdams, H. H. A bacterial cell-cycle regulatory network operating in time and space, Identification of long intergenic repeat sequences associated with DNA methylation sites in Caulobacter crescentus and other alpha-proteobacteria, Fluorescence bleaching reveals asymmetric compartment formation prior to cell division in Caulobacter.
Strains with mutations in one of these genes, flaS, cannot transcribe flagellar structural genes and divide abnormally.
Saurabh, S. n., Perez, A. M., Comerci, C. J., Shapiro, L. n., Moerner, W. E. Dynamic translation regulation in Caulobacter cell cycle control.
Surface-relief dielectric phase masks implement a double-helix response at two wavelengths to distinguish two different fluorescent labels and to quantitatively and precisely localize them relative to each other in 3D. In spite of their small size, bacteria have a remarkably complex internal organization and external architecture.
Surface topology creates crystal defects and boundaries, thereby guiding S-layer assembly. Roberts, R. C., Toochinda, C., Avedissian, M., Baldini, R. L., Gomes, S. L., Shapiro, L. Regulation of a heat shock sigma(32) homolog in Caulobacter crescentus, A cell cycle-regulated bacterial DNA methyltransferase is essential for viability.
View details for DOI 10.1073/pnas.1114444108. The RF-2 binding region is similar to a NifA binding site normally used in the activation of some sigma 54 promoters involved in nitrogen fixation in other bacteria.
S-layer proteins (SLPs) regulate their extracellular self-assembly by crystallizing when exposed to an environmental trigger. Cell Fate Regulation Governed by a Repurposed Bacterial Histidine Kinase. View details for Web of Science ID 000269372600017, View details for PubMedCentralID PMC2737981, View details for Web of Science ID 000207861909276, View details for Web of Science ID 000207857800509.
--Massachusetts Institute of Technology, Dept.
Here, we identify DipM, a putative LytM endopeptidase in Caulobacter crescentus, and show that it plays a critical role in maintaining cell envelope architecture during growth and division. On the basis of these interactions, a trans-acting hierarchy of flagellar and chemotaxis gene expression is proposed.
Quon, K. C., Marczynski, G. T., Shapiro, L. USE OF FLOW-CYTOMETRY TO IDENTIFY A CAULOBACTER 4.5 S RNA TEMPERATURE-SENSITIVE MUTANT DEFECTIVE IN THE CELL-CYCLE, A DEVELOPMENTALLY-REGULATED CHROMOSOMAL ORIGIN OF REPLICATION USES ESSENTIAL TRANSCRIPTION ELEMENTS. B.S.
View details for DOI 10.1073/pnas.0402606101, View details for Web of Science ID 000222278600018, View details for PubMedCentralID PMC438963. View details for Web of Science ID A1996UD48400020, View details for PubMedCentralID PMC177887. View details for Web of Science ID A1993LX92900005, View details for Web of Science ID A1993LF06100004, View details for Web of Science ID A1993LF06100002, View details for Web of Science ID A1993KX96501075, View details for Web of Science ID A1993KX96501060.
A mutant of C. crescentus that fails to synthesize flagellin has been isolated. Nature Communications13, 1585 (2022). Abedi MH#, Yao M#, Mittelstein DR, Bar-Zion A, Swift MB, Lee-Gosselin A, Barturen-Larrea P, Buss MT, Shapiro MG*.
However, 27 GANTC sites remained unmethylated throughout the cell cycle, suggesting that these protected sites could participate in epigenetic regulatory functions. Neither phosphatidylethanolamine nor its precursor phosphatidylserine was detected.
Thus this website will only describe the research activities of Dr. --Massachusetts Institute of Technology, Dept.
Analysis of deletions and base substitutions in the 5' region of the operon established the presence of two functional promoters: a heat shock-inducible promoter, P1, with characteristics of a sigma 32 promoter, and an adjacent sigma 70-like promoter, P2.
View details for DOI 10.1016/j.cell.2021.08.019. 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 fourth heat-shock protein was detected with antibody to the C. crescentus RNA polymerase. A., Deacon, A. M., Shapiro, L. Using Optically Reversible Spatial Mutations to Dissect the Asymmetric Developmental Program of a Bacterium. Using cluster analysis of the resulting set of 12-element vectors for each of these strains, we identified 52 strains with mutations that affected the localization pattern of the three tagged proteins. The Beachy lab studies the function of Hedgehog proteins and other extracellular signals in morphogenesis (pattern formation) and in injury repair and regeneration (pattern maintenance). Rather than being a passive process, it involves rapid movement of parts of the circular chromosome. Epistasis experiments demonstrated that the fliIJ operon is located in class II of the C. crescentus flagellar regulatory hierarchy, suggesting that the gene products act at an early stage in flagellar assembly. Copyright 2023 Mikhail G. Shapiro | Powered by, on Ultrasound-controlled probiotic therapy, on Ultrasensitive imaging of gene expression, International Symposium on Biomolecular Ultrasound and Sonogenetics, Ultrasensitive imaging of gene expression.
The bacterial heat shock proteins DnaK and DnaJ are members of a class of molecular chaperones that are required for a wide variety of cellular functions at normal growth temperatures.
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