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111.
112.
ΦX174 lysis protein E-mediated lysis of Escherichia coli is characterized by a protein E-specific fusion of the inner and outer membrane and formation of a transmembrane tunnel structure. In order to understand the fusion process, the topology of protein E within the envelope complex of E. coli was investigated. Proteinase K protection studies showed that, during the time course of protein E-mediated lysis process, more of the fusion protein E-FXa-streptavidin gradually became accessible to the protease at the cell surface. These observations postulate a conformational change in protein E during induction of the lysis process by movement of the C-terminal end of the protein throughout the envelope complex from the inner side to the outer side spanning the entire pore and fusing the inner and outer membranes at distinct areas. The initiation mechanism for such a conformational change could be the cis–trans isomerization of proline residues within α-helical membrane-spanning segments. Conversion of proline 21, presumed to be in the membrane-embedded α-helix of protein E, to alanine, glycine, serine and valine, respectively, resulted in lysis-negative E mutant proteins. Proteinase K accessibility studies using streptavidin as a reporter fused to the P21G mutant protein showed that the C-terminal part of the fusion protein is not translocated to the outer side of the membrane, suggesting that this proline residue is essential for the correct folding of protein E within the cell wall complex of E. coli . Oligomerization of protein P21G-StrpA was not disturbed. 相似文献
113.
Lioubov I. Tikhonova Igor I. Pottosin Karl-Josef Dietz Gerald Schönknecht 《The Plant journal : for cell and molecular biology》1997,11(5):1059-1070
In contrast to the vacuolar ion channels which are gated open by an increase of cytosolic Ca2+ the vacuolar ion currents at resting cytosolic Ca2+ are poorly explored. Therefore, this study was performed to investigate the properties of the so-called fast-activating vacuolar (FV) current which dominates the electrical characteristics of the tonoplast at physiological free Ca2+ concentrations. Patch—clamp measurements were performed on whole barley ( Hordeum vulgare ) mesophyll vacuoles and on excised tonoplast patches. Single ion channels were identified, which, based on their selectivity, activation kinetics, Ca2+ - and voltage-dependence, carry the whole-vacuole FV current. Reversal potential determinations indicated a K+ overs C− permeability ratio of about 30. Both inward and outward whole-vacuole currents as well as the activity of single FV channels were inhibited by an increase of cytosolic Ca2+ , with a Kd ≈ 6 µM. At physiological vacuolar Ca2+ activities, the FV channel is an outward-rectifying potassium channel. The FV channel was activated in less than a few milliseconds both by negative and positive potential steps, having a minimal activity that is 40 mV negative of the K+ equilibrium potential. It is proposed that transport of K+ through this cation channel controls the electrical potential difference across the tonoplast. 相似文献
114.
115.
Dictyostelium RasG Is Required for Normal Motility and Cytokinesis, But Not Growth 总被引:1,自引:0,他引:1
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Richard I. Tuxworth Janet L. Cheetham Laura M. Machesky George B. Spiegelmann Gerald Weeks Robert H. Insall 《The Journal of cell biology》1997,138(3):605-614
RasG is the most abundant Ras protein in growing Dictyostelium cells and the closest relative of mammalian Ras proteins. We have generated null mutants in which expression of RasG is completely abolished. Unexpectedly, RasG− cells are able to grow at nearly wild-type rates. However, they exhibit defective cell movement and a wide range of defects in the control of the actin cytoskeleton, including a loss of cell polarity, absence of normal lamellipodia, formation of unusual small, punctate polymerized actin structures, and a large number of abnormally long filopodia. Despite their lack of polarity and abnormal cytoskeleton, mutant cells perform normal chemotaxis. However, rasG− cells are unable to perform normal cytokinesis, becoming multinucleate when grown in suspension culture. Taken together, these data suggest a principal role for RasG in coordination of cell movement and control of the cytoskeleton. 相似文献
116.
While globin genes ctt-2β and ctt-9.1 in Chironomus thummi thummi each have a single intron, all of the other insect globin genes reported so far are intronless. We analyzed four globin genes
linked to the two intron-bearing genes in C. th. thummi. Three have a single intron at the same position as ctt-2β and ctt-9.1; the fourth is intronless and lies between intron bearing genes. Finally, in addition to its intron, one gene (ctt-13RT) was recently interrupted by retrotransposition. Phylogenetic analyses show that the six genes in C. th. thummi share common ancestry with five globin genes in the distantly related species C. tentans, and that a 5-gene ancestral cluster predates the divergence of the two species. One gene in the ancestral cluster gave rise
to ctn-ORFB in C. tentans, and duplicated in C. th. thummi to create ctt-11 and ctt-12. From parsimonious calculations of evolutionary distances since speciation, ctt-11, ctt-12, and ctn-ORFB evolved rapidly, while ctn-ORFE in C. tentans evolved slowly compared to other globin genes in the clusters. While these four globins are under selective pressure, we
suggest that most chironomid globin genes were not selected for their unique function. Instead, we propose that high gene
copy number itself was selected because conditions favored organisms that could synthesize more hemoglobin. High gene copy
number selection to produce more of a useful product may be the basis of forming multigene families, all of whose members
initially accumulate neutral substitutions while retaining essential function. Maintenance of a large family of globin genes
not only ensured high levels of hemoglobin production, but may have facilitated the extensive divergence of chironomids into
as many as 5000 species.
Received: 31 December 1996 / Accepted: 16 May 1997 相似文献
117.
Brief overview of control of genetic expression by antisense oligonucleotides and in vivo applications 总被引:7,自引:0,他引:7
Gerald Zon 《Molecular neurobiology》1995,10(2-3):219-229
Over the past several years, the use of synthetic oligonucleotides and functional analogs thereof as a possibly general means
of controlling genetic expression has received widespread attention. Following a brief overview of some of the basic principles
and strategies for this approach, attention is focused here on summarizing some recent reports of in vitro and, in particular,
in vivo investigations in various animal models using phosphorothioate analogs of 2′-deoxyoligo-nucleotides. In view of these
findings, which include studies related to neurobiology, this field should find significant utility in applications of the
antisense method for controlling genetic expression. 相似文献
118.
A stochastic approach is utilized to develop a model equation capable of describing the time course of germination in a sample
of bacterial spores. The time required by a spore to complete the change characteristic of germination consists of an initial
interval of no change followed immediately by the duration of the change itself. The experimental basis of the proposed model
is the observation that each of these time intervals is distributed over a range of values in a spore sample. Mixed continuous
and discrete probabilities are employed in arriving at an average single-spore germination curve which, to a different scale,
describes the sample in time. 相似文献
119.
120.
Michael J. Barber Andrew S. Zektzer Gerald M. Rosen Helen A. Demos Elmer J. Rauckman 《生物化学与生物物理学报:生物膜》1984,776(1):159-168
Hepatic microsomal membranes, prepared under various conditions that yield either ‘intact’ or ‘disrupted’ microsomal vesicles, have been labeled via the sulfhydryl groups of intrinsic membrane proteins using nitroxide analogs of . Electron paramagnetic resonance spectra revealed the presence of two dominant classes of bound label corresponding to differing degrees of immobilization, the ratio of which were quantitated using a parameter designated the ‘’ ratio. For latent microsomes, the value of this parameter was determined to be and was influenced by factors such as label/protein ratio, incubation period, nitroxide structure, temperature and pH. The ratio was also sensitive to the degree of membrane integrity as revealed by the latency of mannose 6-phosphate activity of glucose-6-phosphohydrolase. In addition, membrane disruption resulted in a corresponding decrease in the order parameter for nitroxide-labeled fatty acids intercalated within the lipid bilayer. The ratio was observed to be dependent upon the method of microsome preparation yielding values of for ‘hypertonically disrupted’ vesicles and for ‘mechanically disrupted’ vesicles. Microsomal marker enzymes such as cytochrome and FAD-containing monooxygenase retained significant levels of functionally following nitroxide incorporation. 相似文献