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Insertional inactivation of genes encoding components of the sodium-type flagellar motor and switch of Vibrio parahaemolyticus 下载免费PDF全文
Vibrio parahaemolyticus possesses two types of flagella, polar and lateral, powered by distinct energy sources, which are derived from the sodium and proton motive forces, respectively. Although proton-powered flagella in Escherichia coli and Salmonella enterica serovar Typhimurium have been extensively studied, the mechanism of torque generation is still not understood. Molecular knowledge of the structure of the sodium-driven motor is only now being developed. In this work, we identify the switch components, FliG, FliM, and FliN, of the sodium-type motor. This brings the total number of genes identified as pertinent to polar motor function to seven. Both FliM and FliN possess charged domains not found in proton-type homologs; however, they can interact with the proton-type motor of E. coli to a limited extent. Residues known to be critical for torque generation in the proton-type motor are conserved in the sodium-type motor, suggesting a common mechanism for energy transfer at the rotor-stator interface regardless of the driving force powering rotation. Mutants representing a complete panel of insertionally inactivated switch and motor genes were constructed. All of these mutants were defective in sodium-driven swimming motility. Alkaline phosphatase could be fused to the C termini of MotB and MotY without abolishing motility, whereas deletion of the unusual, highly charged C-terminal domain of FliM disrupted motor function. All of the mutants retained proton-driven, lateral motility over surfaces. Thus, although central chemotaxis genes are shared by the polar and lateral systems, genes encoding the switch components, as well as the motor genes, are distinct for each motility system. 相似文献
94.
Evolutionary conservation of methyl-accepting chemotaxis protein location in Bacteria and Archaea 下载免费PDF全文
Gestwicki JE Lamanna AC Harshey RM McCarter LL Kiessling LL Adler J 《Journal of bacteriology》2000,182(22):6499-6502
The methyl-accepting chemotaxis proteins (MCPs) are concentrated at the cell poles in an evolutionarily diverse panel of bacteria and an archeon. In elongated cells, the MCPs are located both at the poles and at regions along the length of the cells. Together, these results suggest that MCP location is evolutionarily conserved. 相似文献
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Rampant horizontal transfer and duplication of rubisco genes in eubacteria and plastids 总被引:16,自引:2,他引:14
Previous work has shown that molecular phylogenies of plastids,
cyanobacteria, and proteobacteria based on the rubisco (ribulose-1,5-
bisphosphate carboxylase/oxygenase) genes rbcL and rbcS are incongruent
with molecular phylogenies based on other genes and are also incompatible
with structural and biochemical information. Although it has been much
speculated that this is the consequence of a single horizontal gene
transfer (of a proteobacterial or mitochondrial rubisco operon into
plastids of rhodophytic and chromophytic algae), neither this hypothesis
nor the alternative hypothesis of ancient gene duplication have been
examined in detail. We have conducted phylogenetic analyses of all
available bacterial rbcL sequences, and representative plastid sequences,
in order to explore these alternative hypothesis and fully examine the
complexity of rubisco gene evolution. The rbcL phylogeny reveals a
surprising number of gene relationships that are fundamentally incongruent
with organismal relationships as inferred from multiple lines of other
molecular evidence. On the order of six horizontal gene transfers are
implied by the form I (L8S8) rbcL phylogeny, two between cyanobacteria and
proteobacteria, one between proteobacteria and plastids, and three within
proteobacteria. Alternatively, a single ancient duplication of the form I
rubisco operon, followed by repeated and pervasive differential loss of one
operon or the other, would account for much of this incongruity. In all
probability, the rubisco operon has undergone multiple events of both
horizontal gene transfer and gene duplication in different lineages.
相似文献
98.
Nonparametric regression in the presence of measurement error 总被引:4,自引:0,他引:4
99.
Qingchun Zhang Matthew R. Schenauer John D. McCarter Gregory C. Flynn 《The Journal of biological chemistry》2013,288(23):16371-16382
During either production or storage, the LC214-HC220 disulfide in therapeutic antibodies can convert to a thioether bond. Here we report that a thioether forms at the same position on antibodies in vivo. An IgG1κ therapeutic antibody dosed in humans formed a thioether at this position at a rate of about 0.1%/day while circulating in blood. Thioether modifications were also found at this position in endogenous antibodies isolated from healthy human subjects, at levels consistent with this conversion rate. For both endogenous antibodies and recombinant antibodies studied in vivo, thioether conversion rates were faster for IgG1 antibodies containing λ light chains than those containing κ light chains. These light chain reaction rate differences were replicated in vitro. Additional mechanistic studies showed that base-catalyzed thioether formation through the light chain dehydrogenation was more preferred on antibodies with λ light chains, which may help explain the observed reaction rate differences. 相似文献
100.
Sarah D. McCarter Debra L. Johnson Khameeka N. Kitt Carolyn Donohue Alison Adams Jean M. Wilson 《Traffic (Copenhagen, Denmark)》2010,11(6):856-866
The establishment of tight junctions and cell polarity is an essential process in all epithelia. Endotubin is an integral membrane protein found in apical endosomes of developing epithelia when tight junctions and epithelial polarity first arise. We found that the disruption of endotubin function in cells in culture by siRNA or overexpression of the C‐terminal cytoplasmic domain of endotubin causes defects in organization and function of tight junctions. We observe defects in localization of tight junction proteins, reduced transepithelial resistance, increased lanthanum penetration between cells and reduced ability of cells to form cysts in three‐dimensional culture. In addition, in cells overexpressing the C‐terminal domain of endotubin, we observe a delay in re‐establishing the normal distribution of endosomes after calcium switch. These results suggest that endotubin regulates trafficking of polarity proteins and tight junction components out of the endosomal compartment, thereby providing a critical link between a resident protein of apical endosomes and tight junctions. 相似文献