Immunoassays of colonies lysed in situ either by lambda prophage induction or by biochemical means afford a much higher level of sensitivity than the plaque assay probably adequate to detect the production of a few molecules of protein per cell. Affiliations: Bioengineering, Biochemistry, Neurobiology View details for Web of Science ID 000430563200493, View details for Web of Science ID 000430450000249, View details for Web of Science ID 000430563200402, View details for Web of Science ID 000430450000508, View details for Web of Science ID 000430563300065, View details for Web of Science ID 000430439600509. John Heath, SURF Scholar 2018-20 PhD at Stanford Cell type- and compartment-specific localization of the CtrA global transcriptional regulator is essential for proper cell-cycle progression, and subcellular localization of key chromosome partitioning proteins is correlated with proper nucleoid segregation. Collaboration: In-vivo Drug Evaluation, University of Colorado and Health Sciences Center
View details for Web of Science ID A1992KB97700015. Ph.D. Student, Bioengineering We also study the normal roles of such signals in stem-cell physiology and their abnormal roles in the formation and expansion of cancer stem cells. Undergraduate Researcher 2019- Quantitative Multicolor Subdiffraction Imaging of Bacterial Protein Ultrastructures in Three Dimensions. These basal bodies have five rings threaded on a rod. Although these strains exhibit different morphologies and DNA content, the replication of both strains in macrophages is attenuated. We study the physiology, modulation and functional role of these channels in heterologous expression systems, dissociated neurons and in brain slice from the hippocampus and other regions. Our laboratory is using genetic mapping, comparative sequence analysis, and functional tests to identify the genomic basis of classic evolutionary traits in vertebrates. Differential translation of operon-encoded genes facilitates precise cell cycle-timing for the dynamic assembly of multiprotein complexes, such as the flagellum and the stalk and the correct positioning of regulatory proteins to specific cell poles. View details for DOI 10.1073/pnas.1906119116, View details for DOI 10.1016/j.bpj.2018.11.881, View details for Web of Science ID 000460779800789, View details for DOI 10.1016/j.bpj.2018.11.2446, View details for Web of Science ID 000460779802279, View details for DOI 10.1016/j.bpj.2018.11.056, View details for Web of Science ID 000460779800028, View details for DOI 10.1016/j.bpj.2018.11.1077, View details for Web of Science ID 000460779800965, View details for DOI 10.1016/j.bpj.2018.11.2475, View details for Web of Science ID 000460779802305. This C. crescentus secA mutant has allowed the identification of morphogenetic events in the swarmer-to-stalked cell transition that require SecA-dependent protein translocation. Out of 2726 TSSs, 586 were shown to be cell cycle-regulated and we identified 529 binding sites for the cell cycle master regulators. The ubiquitous DnaA protein is a major regulator of all three bacterial origins. article. Furthermore, we introduce a newly constructed set of integrating and replicating shuttle vectors that considerably facilitate cell biological and physiological studies in Caulobacter. Viollier, P. H., Sternheim, N., Shapiro, L. A signal transduction protein cues proteolytic events critical to Caulobacter cell cycle progression, DNA methylation affects the cell cycle transcription of the CtrA global regulator in Caulobacter, A dynamically localized histidine kinase controls the asymmetric distribution of polar pili proteins, Biomolecular screening with encoded porous-silicon photonic crystals. We demonstrate here that the expression of the Escherichia coli chemoreceptor gene tsr, with 2.6 kilobases of its upstream sequence, is temporally controlled in Caulobacter crescentus. The specific defects responsible for the lack of a chemotactic response have not been determined for the other identified che genes. Here, we show that the cytoplasmic response regulator, DivK, exhibits a dynamic, cyclical localization that culminates in asymmetric distribution of DivK within the two cell types that are characteristic of the Caulobacter cell cycle; DivK is dispersed throughout the cytoplasm of the progeny swarmer cell and is localized to the pole of the stalked cell. Here we show that the MS-ring monomer FliF, a central motor component that anchors the flagellum in the cell membrane, is synthesized only in the predivisional cell and is integrated into the membrane at the incipient swarmer cell pole, where it initiates flagellar assembly. 169:1493-1498, 1987). Analysis of the cloned and sequenced dnaK gene has shown that the deduced amino acid sequence could encode a protein of 67.6 kilodaltons that is 68% identical to the DnaK protein of Escherichia coli and 49% identical to the Drosophila and human hsp70 protein family. View details for DOI 10.1128/mBio.03020-20. Caulobacter crescentus initiates a single round of DNA replication during each cell cycle. Homologs of CcrM are widespread in the alpha subdivision of the Proteobacteria, suggesting that methylation at GANTC sites may have important functions in other members of this diverse group as well. B.S. A Bacterial Toxin Perturbs Intracellular Amino Acid Balance To Induce Persistence. The contribution of each promoter for genes transcribed from multiple promoters is identified. These results suggest that subcellular localization of a prokaryotic protein involves interaction of specific regions of the protein with unique cell sites that contain either localized binding proteins or a specific secretory apparatus. During the swarmer-to-stalked transition, PodJS must be degraded to preserve asymmetry in the next cell cycle. We have identified a periplasmic protease (PerP) that initiates the proteolytic sequence by truncating PodJ(L) to a form with altered activity (PodJ(S)). The phage is composed of 57% DNA. Biology, Dongguk University Ph.D. Student, Neurobiology (co-advised with David Anderson) Each cell division in Caulobacter crescentus is asymmetric, yielding a swarmer cell with several polar pili and a non-piliated stalked cell. The biogenesis of the polar flagellum in Caulobacter crescentus is limited to a specific time in the cell cycle and to a specific site on the cell. Under these conditions, the cell enters a quiescent state referred to as dormancy or persistence. Tome Biosciences, Prof. JrmeLacroix View details for Web of Science ID 000167833700095, View details for PubMedCentralID PMC31192. Cytokinesis in Gram-negative bacteria is mediated by a multiprotein machine (the divisome) that invaginates and remodels the inner membrane, peptidoglycan and outer membrane. Shapiro received his bachelors from Brooklyn College, before earning his PhD from Purdue University. Stanford University; Joe Shapiro, University of California Berkeley; Deadline for paper submission. View details for Web of Science ID A1994PM98800011. This Tn5 derivative also contained the intact tetracycline resistance-encoding region of the transposon Tn10. Maria Paulene Abundo The enzyme acts processively during methylation of specific DNA sequences, indicating a preferred order of product release in which S-adenosylhomocysteine is released from enzyme before fully methylated DNA. These compounds were identified by screening for inhibitors against Caulobacter crescentus CcrM, an essential DNA methyltransferase from gram negative alpha-proteobacteria. Physics, expected 2023 Biological Engineering, MIT Furthermore, carbon source starvation results in accumulation of the cells at the predivisional stage. 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. For questions or comments, contact the SLAC Office of Communications at communications@slac.stanford.edu. The genes involved in the biogenesis of the flagellum and the chemotaxis machinery are temporally regulated during the Caulobacter crescentus cell cycle. Degradation of the CtrA response regulator normally coincides with initiation of DNA replication, but in strains lacking SsrA activity there is a 40-min delay between the degradation of CtrA and replication initiation. View details for Web of Science ID 000251336000004, View details for PubMedCentralID PMC2175322. View details for Web of Science ID A1995QB30700010, View details for PubMedCentralID PMC176597, View details for Web of Science ID A1995BG35H00001. The synthesis of the major C. crescentus RNA polymerase sigma factor was not induced by heat shock. BLP Fellow Elin Kang, SURF Scholar 2021 BS Chemistry, Caltech 2023 (exp) Biochemistry, Jagiellonian University The hybridization method used permits the detection of sequences partially homologous to the elements. We determined the cellular positions of the origins of the replicons in the alpha proteobacteria Agrobacterium tumefaciens and Sinorhizobium meliloti and found that they are located at the poles of the cells. Nucleoid morphology was also abnormal. The rest of the filament (region V) is made up predominantly, if not completely, of the 25 x 10(3) Mr flagellin. Several temporally controlled flagellar genes in Caulobacter crescentus require a sigma 54 promoter and upstream sites for transcription activation. The genes in this cluster form an operon whose expression is controlled temporally. Michael Yao, SURF Scholar 2017-21 (Housner Prize) MD-PhD at University of Pennsylvania 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. They have found a single molecular event present in all cancers studied to date that protects them from macrophages of the innate immune system. There have been two sharp demarcations in my life in science: the transition from fine arts to chemistry, which happened early in my career, and the move from New York to Stanford University, which initiated an ongoing collaboration with the physicist Harley McAdams. Interested applicants should visit https://facultypositions.stanford.edu/en-us/job/493432 for our full ad and more information about how to apply. Cedars-Sinai Medical Center, Dr. Mohamad Abedi View details for DOI 10.1038/sj.emboj.7600935, View details for Web of Science ID 000234952500011, View details for PubMedCentralID PMC1383518. This segregation does not depend on sequences within the mRNA, but on the upstream regulatory region. x@caltech.edu, x=wcoleman, George Daghlian 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. Thanbichler, M., Iniesta, A. Ph.D. 1972 Purdue University
Dibutyryl cyclic adenosine 3',5'-monophosphate (AMP) was shown to stimulate expression of the inducible enzymes and, thus, the initiation of the cell cycle. Thus, the sequential degradation of PodJ appears to involve regulated intramembrane proteolysis (Rip) by MmpA. Active segregation by a mitotic machinery appears to be common; however, the mode of chromosome segregation varies significantly from species to species. The C-terminal crystallization domain forms the physiological 2-dimensional (2D) crystal lattice, but full-length protein crystallizes multiple orders of magnitude faster due to the N-terminal nucleation domain. During development of the symbiotic soil bacterium Sinorhizobium meliloti into nitrogen-fixing bacteroids, DNA replication and cell division cease and the cells undergo profound metabolic and morphological changes. The CtrA master transcriptional regulator is a central control element in Caulobacter cell cycle progression and polar morphogenesis. The transcription of the che operon occurred at a defined time in the cell cycle, prior to cell division. View details for Web of Science ID A1996TR39200015, View details for PubMedCentralID PMC177711. The ribonucleic acid (RNA) bacteriophage phiCb5, which specifically infects only one form of the dimorphic stalked bacterium Caulobacter crescentus, has been obtained in high yield. Bryan, R., CHAMPER, R., Gomes, S., Ely, B., Shapiro, L. GENERAL NONCHEMOTACTIC MUTANTS OF CAULOBACTER-CRESCENTUS. In a mutant strain that failed to assemble a flagellum, the 29K flagellin still segregated to the presumptive swarmer cell, demonstrating that positioning of the protein is independent of filament assembly. The flbN-encoded protein has 30.8% identity with the protein encoded by the Salmonella typhimurium basal body L-ring gene, flgH. View details for Web of Science ID A1989R820200026. James Boswell Postdoctoral Scholar This involves moving toward food, adapting to environmental extremes, and, in many cases, entering and exploiting a eukaryotic host. Iniesta, A. Replisome assembly occurs at the chromosomal origin located at the stalked cell pole, coincident with the initiation of DNA replication. Chemical Engineering View details for Web of Science ID A1980JS45300028, View details for Web of Science ID A1980LV23700014, View details for Web of Science ID A1980JN48100014. Only the stalked cell progeny is able to replicate its chromosome, and the swarmer cell progeny must differentiate into a stalked cell before it too can replicate its chromosome. Postdoctoral Scholar, 2017-2021 DivL uniquely contains a tyrosine at the histidine phosphorylation site, and can achieve these regulatory functions in vivo without kinase activity. All four proteins were synthesized in the predivisional cell, but the progeny showed cell type-specific bias in the level of enhanced synthesis after heat shock. Postdoctoral Scholar (co-advised with Mory Gharib) Menlo Park, Calif. The Department of Energys SLAC National Accelerator Laboratory and Stanford University today announced the launch of a new joint battery Using these motifs, we predict coregulated genes. Regulated proteolysis of GcrA contributes to the cell cycle variations in GcrA abundance. These data suggest a more prevalent use of the Shine-Dalgarno sequence for ribosome pausing rather than translation initiation in C. crescentus. We conclude that in bacteria, as in yeast, (i) genes involved in a given cell function are activated at the time of execution of that function, (ii) genes encoding proteins that function in complexes are coexpressed, and (iii) temporal cascades of gene expression control multiprotein structure biogenesis. Chen, J. C., Hottes, A. K., McAdams, H. H., McGrath, P. T., Viollier, P. H., Shapiro, L. Two independent spiral structures control cell shape in Caulobacter. The N-terminal proteolytic determinant is predicted to reside on the surface of the receiver domain in beta-sheet 2 and alpha-helix 2. After graduating he worked for the NASA Marshall Space Flight Center developing non-destructive evaluation techniques for applications related to the US space program. Our major publications have shown the PIP2 sensitivity of both types of channels, the mechanisms and structural determinants of receptor-mediated suppression of IM and ICa, the modulation of KCNQ channels by calmodulin, A-kinase anchoring proteins (AKAPs) and Src kinase, the roles of M channels in airway smooth muscle, and in sensory neurons. Temporal control of DNA methylation has been shown to be critical for normal development in the dimorphic Caulobacter life cycle. Focused ultrasound excites cortical neurons via mechanosensitive calcium accumulation and ion channel amplification. A null mutation in the Caulobacter crescentus smc gene is conditionally lethal and causes a cell cycle arrest at the predivisional cell stage. Hillson, N. J., Hu, P., Andersen, G. L., Shapiro, L. A comprehensive set of plasmids for vanillate- and xylose-inducible gene expression in Caulobacter crescentus. x@caltech.edu, x=hao.k.shen, Shirin Shivaei Transcription of flagellar genes in Caulobacter crecentus is programmed to occur during the predivisional stage of the cell cycle. DEOXYRIBONUCLEIC-ACID SEQUENCE HOMOLOGIES AMONG BACTERIAL INSERTION SEQUENCE ELEMENTS AND GENOMES OF VARIOUS ORGANISMS, CELL-CYCLE-ASSOCIATED REARRANGEMENT OF INVERTED REPEAT DNA-SEQUENCES. The resulting models illustrate that the genome is ellipsoidal with periodically arranged arms. x@caltech.edu, x=blling, Ann Liu Currently: Assistant Professor of Bioengineering Analysis of this organism is complicated by a limited selection of tools for genetic manipulation and inducible gene expression. View details for Web of Science ID A1972M472200040, View details for Web of Science ID A1971J193000005. The cellular location of a derivative of the RK2 plasmid is distinct from that of the alpha proteobacterium genomic replicon origins but is conserved across bacteria. Ph.D. Student, Biochemistry & Molecular Biophysics (co-ad. View details for Web of Science ID A1995RM70200022. The uranium reporter construct was effective for discriminating contaminated groundwater samples (4.2 microM uranium) from uncontaminated groundwater samples (<0.1 microM uranium) collected at the Oak Ridge Field Research Center. We determined the plasmid copy number in both progeny cell types, and determined that plasmids partitioned equally to the stalked and swarmer cells. A single flagellum is assembled at the swarmer pole of the predivisional cell and is released later in the cell cycle. Although FDA is exercising enforcement discretion of premarket review and other regulations for laboratory-developed tests in the US, certification of the laboratory is required under CLIA to ensure the quality and validity of the tests. Hear Gigis story. Postdoctoral Scholar Because regulatory proteins are among those that reside at specific cellular sites, it is now necessary to consider three-dimensional organization when describing the genetic networks that control bacterial cells. Furthermore, the four heat-shock proteins synthesized in the predivisional cell were partitioned in a specific manner upon cell division. Our tests are clinically validated in over 100, Limited Noninvasive Prenatal Testing (NIPT), Schedule Session with Patient Coordinator, Order Tests and Track Status on NateraConnect, Notice of Data Collection for California Residents. CrfA functions to stabilize the CC3461 transcript. In particular, the distribution of HU, which is the most abundant NAP, has received little attention. In nature, this essential process occurs in cells that live in fluctuating environments. Using in vivo and in vitro analyses of dynamic polar protein complex formation, we show that a polymeric cell polarity protein, SpmX, serves as a direct bridge between the PopZ polymeric network and the cell fate-directing DivJ histidine kinase. x@caltech.edu, x=syoo1984, Research Staff Our correlative approach reveals that SpmX localizes along one side of the cell pole and its extent closely matches that of the PopZ region. Our algorithm takes into account information about a beam that is normally discarded and uses that information to paint a more detailed picture of the beam.. WebLucy Shapiro's Profile | Stanford Profiles Email Profile Academic Appointments Professor, Developmental Biology Member, Bio-X Faculty Fellow, Sarafan ChEM-H Administrative No extrachromosomal elements were found in spite of systematic attempts to detect their presence. Ramya Deshpande, SURF Scholar 2019-20 PhD at Harvard There are many instances of differential polar functions; among these is the preferential use of old poles when attaching to host cells as in the interaction of Bradyrhizobium with plant root hairs (3) or the polar pili-mediated attachment of the Pseudomonas aeruginosa pathogen to tracheal epithelia (4). B.S. We report that SciP, a helix-turn-helix transcription factor, is an essential component of this circuit. For questions or comments, contact the SLAC Office of Communications atcommunications@slac.stanford.edu. Ph.D. Biomedical Engineering, University of Minnesota Thus, the Fix network is a conserved sensory/signaling module whose transcriptional output has been adapted to the unique physiologies of C. crescentus and the nitrogen-fixing rhizobia. View details for Web of Science ID 000170907900034. 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 10.1073/pnas.0402606101, View details for Web of Science ID 000222278600018, View details for PubMedCentralID PMC438963. CtrA activity in the cell cycle is controlled both transcriptionally and by phosphorylation. Barnett, M. J., Hung, D. Y., Reisenauer, A., Shapiro, L., Long, S. R. The CcrM DNA methyltransferase of Agrobacterium tumefaciens is essential, and its activity is cell cycle regulated. View details for Web of Science ID A1992HU93800030. Each Caulobacter cell cycle involves differentiation and an asymmetric cell division driven by a cyclical regulatory circuit comprised of four transcription factors (TFs) and a DNA methyltransferase. They also allow an investigation of the regulatory signals that modulate the asymmetric distribution of proteins and their subsequent cell type-specific expression in the initial stages of a developmental program. View details for Web of Science ID A1989AP19100046. Expression of the ccrM gene was found to be restricted to the portion of the cell cycle immediately prior to cell division. The 92,000 Mr Lon homolog and the 37,000 Mr RNA polymerase subunit were preferentially synthesized in the stalked cell, whereas the synthesis of the 62,000 Mr GroEL homolog was enhanced in the progeny swarmer cell. This structure is absent at the flagellar pole but not in the stalks of flbT mutant predivisional cells. Three global regulatory proteins cycle out of phase with one another and drive cell cycle progression by directly controlling the expression of 200 cell-cycle-regulated genes. Extensive mutational analysis of the promoter region from -42 to -5 identified functionally important nucleotides at -36 and -35, between -29 and -22, and at -12, which correlates well with sequences conserved between fliLM and the analogous regions of two other Class II flagellar operons. The algorithm pairs machine-learning techniques with beam physics equations to avoid massive data crunching. B.A.Sc. Mathematical and Computational Biology, Harvey Mudd College We are interested in how Arctic species and populations responded to environmental and habitat change throughout the Pleistocene, and what role ecology, natural history, climate and community-level dynamics played in the M.S. Adduct formation is mediated through the boron atom of AN2690 and the 2'- and 3'-oxygen atoms of tRNA's3'-terminal adenosine. CHAMPER, R., Bryan, R., Gomes, S. L., Purucker, M., Shapiro, L. ANALYSIS OF THE PLEIOTROPIC REGULATION OF FLAGELLAR AND CHEMOTAXIS GENE-EXPRESSION IN CAULOBACTER-CRESCENTUS BY USING PLASMID COMPLEMENTATION. Furthermore, the FtsK N terminus is required to either assemble or maintain FtsZ rings at the division plane. The ffs36 phenotype results from a single base change in one of the non-conserved stems of the mature RNA, and is completely rescued by a compensating mutation in the opposite strand, providing confirmation of the predicted secondary structure of the 4.5 S RNA. Similarly, hooks with attached rods were shed from nonflagellate mutants, and these structures also lacked the basal rings. S1 mapping and primer extension experiments showed that transcription initiated at two sites 5' to the dnaK coding sequence. In particular, super-resolution microscopy methods overcome the diffraction limit to observe nanoscale cellular structures with unprecedented detail, and single-molecule tracking provides precise dynamic information about the motions of labeled proteins and oligonucleotides. We perform cryogenic super-resolution experiments in situ, labeling PopZ, a protein known to assemble into a microdomain at the poles of the model bacterium Caulobacter crescentus. View details for Web of Science ID 000080842200015, View details for PubMedCentralID PMC21969, View details for Web of Science ID 000080527100001, View details for PubMedCentralID PMC34200. View details for Web of Science ID A1993MG71100001. Fluorescence microscopy is a sensitive tool for this purpose. czhang8@illinois.edu
View details for DOI 10.1101/cshperspect.a000349, View details for Web of Science ID 000279881400003, View details for PubMedCentralID PMC2828278. Small non-coding RNAs (sRNAs) are active in many bacterial cell functions, including regulation of the cell's response to environmental challenges. 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. SLAC is a vibrant multiprogram laboratory that explores how the universe works at the biggest, smallest and fastest scales and invents powerful tools used by scientists around the globe. A localized adaptor protein performs distinct functions at the Caulobacter cell poles. Gloria Ha, SURF Scholar 2015 PhD at Harvard The Office of Science is the single largest supporter of basic research in the physical sciences in the United States and is working to address some of the most pressing challenges of our time. article | press. Fluorescence microscopy can potentially be used to reveal this information when specific labels, known as fluorescent biosensors, are used, but there has been minimal use of such biosensors in cryo-CLEM to date. Sequence comparison of the fliL transcription start site with those of other class I genes, flaS and flaO, revealed a highly conserved 29-bp sequence in a potential promoter region that differs from sigma 70, sigma 54, sigma 32, and sigma 28 promoter sequences, suggesting that at least three class I genes share a unique 5' regulatory region. PleA is also required for the assembly of substructures of the flagellar basal body hook complex that are located in or traverse the peptidoglycan layer. Group Affiliate, Senior Professional Staff at Johns Hopkins Applied Physics Laboratory. C. crescentus DNA carrying the Tn5-flaE region and adjacent sequences was cloned into pBR325 and selected by transposon-encoded kanamycin resistance. Because DivL and CckA accumulate at the same cell pole after the initiation of DNA replication and were found to interact in vivo, we propose that DivL recruits CckA to the pole, thereby promoting its autophosphorylation and activity. B.A., Physics, University of Chicago, 1984. Both promoters were also expressed constitutively throughout the cell cycle under physiological conditions. Herrmann, J., Comerci, C. J., Jabbarpour, F., Shapiro, L., Moerner, W. E., Wakatsuki, S. Dissection of Protein Function Within a Bacterial Biomolecular Condensate by In Vitro Reconstitution. The master CtrA response regulator functions in Caulobacter to repress replication initiation in different phases of the cell cycle. Including the physics of particle beam dynamics with the experimental data allowed the researchers to accurately reconstruct fine details of the beam using only 10 data points a task that might take up to 10,000 data points for some machine learning models that dont include a model of beam physics. During his time with us, he searched for a "first principle project" that defines life by In Caulobacter crescentus the biosynthesis and assembly of this structure is under temporal and spatial control. Postdoctoral Scholar, 2014-16 Here we demonstrate that the Caulobacter PopZ scaffold creates an organizing center at the cell pole that actively regulates polar centromere transport by the ParA partition system. View details for Web of Science ID 000294537800007, View details for PubMedCentralID PMC3202797. These promoters, as well as those for several other genes encoding DNA replication proteins that are induced at the same time in the cell cycle, share two sequence motifs, suggesting that they represent a family whose transcription is coordinately regulated. Two-component signal transduction proteins are known to play a significant role in cell cycle progression. View details for Web of Science ID 000079843900013. Complementarity between a region of CrfA and the terminal region of the CC3461 5'-untranslated region (5'-UTR) and also the behavior of a deletion of this region and a site-specific base substitution and a 3-base deletion in the CrfA complementary sequence suggest that CrfA binds to a stem-loop structure upstream of the CC3461 Shine-Dalgarno sequence and stabilizes the transcript.
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