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1.
The processing of secretory preproteins by signal peptidases (SPases) is essential for cell viability. As previously shown for Bacillus subtilis, only certain SPases of organisms containing multiple paralogous SPases are essential. This allows a distinction between SPases that are of major and minor importance for cell viability. Notably, the functional difference between major and minor SPases is not reflected clearly in sequence alignments. Here, we have successfully used molecular phylogeny to predict major and minor SPases. The results were verified with SPases from various bacilli. As predicted, the latter enzymes behaved as major or minor SPases when expressed in B. subtilis. Strikingly, molecular modeling indicated that the active site geometry is not a critical parameter for the classification of major and minor Bacillus SPases. Even though the substrate binding site of the minor SPase SipV is smaller than that of other known SPases, SipV could be converted into a major SPase without changing this site. Instead, replacement of amino-terminal residues of SipV with corresponding residues of the major SPase SipS was sufficient for conversion of SipV into a major SPase. This suggests that differences between major and minor SPases are based on activities other than substrate cleavage site selection.  相似文献   
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Summary The human -crystallin genes constitute a multigene family whose members are only expressed in the eye lens. The chromosomal location of these sequences has been determined by screening a panel of human/rodent hybrid cell lines containing overlapping subsets of human chromosomes for the presence of human -crystallin sequences. By correlating these genomic hybridization data with the chromosomal constitution of the somatic cell hybrids, all human -crystallin sequences could be assigned to chromosome 2. The use of human/hamster cell hybrids derived from human Burkitt lymphoma cells carrying a reciprocal translocation between human chromosomes 2 and 8, allowed a further localization of the sequences to the region 2p12-qter.  相似文献   
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Zusammenfassung An Hand bekannter Daten wird auf aerodynamischer Grundlage eine Theorie des Ruderfluges der Vögel aufgebaut, deren Wesentliches ist, daß fortwährend, also sowohl beim Aufschlag wie beim Niederschlag, ein Vortrieb vom Vogel geleistet wird.Beim Niederschlag entstehen Auftrieb und Vortrieb, beim Aufschlag Abtrieb und Vortrieb.Wie beim Tragflächenflugzeug ist der Vortrieb das erst Nötige, der Auftrieb folgt dann von selbst. Der Vogel muß aber beim Aufschlag, um einen Vortrieb leisten zu können, einen Abtrieb in Kauf nehmen. Es wird an einem Beispiel gezeigt, daß die dadurch auftretenden Hebungen und Senkungen bei der Taube so gering sind, daß sie dem Auge wohl meistens entgehen werden.  相似文献   
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Activity of the Tat machinery for protein transport across the inner membrane of Escherichia coli and the chloroplast thylakoidal membrane requires the presence of three membrane proteins: TatA, TatB and TatC. Here, we show that the Tat machinery of the Gram-positive bacterium Bacillus subtilis is very different because it contains at least two minimal Tat translocases, each composed of one specific TatA and one specific TatC component. A third, TatB-like component is apparently not required. This implies that TatA proteins of B. subtilis perform the functions of both TatA and TatB of E. coli and thylakoids. Notably, the two B. subtilis translocases named TatAdCd and TatAyCy both function as individual, substrate-specific translocases for the twin-arginine preproteins PhoD and YwbN, respectively. Importantly, these minimal TatAC translocases of B. subtilis are representative for the Tat machinery of the vast majority of Gram-positive bacteria, Streptomycetes being the only known exception with TatABC translocases.  相似文献   
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Soluble forms of Bacillus signal peptidases which lack their unique amino-terminal membrane anchor are prone to degradation, which precludes their high-level production in the cytoplasm of Escherichia coli. Here, we show that the degradation of soluble forms of the Bacillus signal peptidase SipS is largely due to self-cleavage. First, catalytically inactive soluble forms of this signal peptidase were not prone to degradation; in fact, these mutant proteins were produced at very high levels in E. coli. Second, the purified active soluble form of SipS displayed self-cleavage in vitro. Third, as determined by N-terminal sequencing, at least one of the sites of self-cleavage (between Ser15 and Met16 of the truncated enzyme) strongly resembles a typical signal peptidase cleavage site. Self-cleavage at the latter position results in complete inactivation of the enzyme, as Ser15 forms a catalytic dyad with Lys55. Ironically, self-cleavage between Ser15 and Met16 cannot be prevented by mutagenesis of Gly13 and Ser15, which conform to the -1, -3 rule for signal peptidase recognition, because these residues are critical for signal peptidase activity.  相似文献   
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Abstract. 1. The endangered butterfly Phengaris alcon exists in two ecotypes (P. ‘alcon’ and P. ‘rebeli’), which inhabit contrasting biotopes (wet and warm/dry grasslands respectively) and use different larval food plants. The initially flower‐bud‐feeding caterpillars complete their development as social parasites of Myrmica ants, and the specificity of these relationships shows geographical variation. 2. We studied the genetic structure of 16 populations (365 individuals) of both ecotypes in eastern Europe, sampling P. ‘rebeli’ in two disjunct areas in Lithuania and southern Poland, and P. ‘alcon’ on Polish localities between them. We analysed the cytochrome c oxidase subunit I (COI) mitochondrial gene, the EF1‐α nuclear gene and five polymorphic microsatellite loci. 3. All individuals shared an identical COI haplotype, which we hypothesise may be linked to a selective sweep associated with the presence of the Wolbachia B strain in all populations. 4. For nuclear markers, we did not find a clear pattern reflecting division into two putative ecotypes. However, ecotypes differed significantly in their genetic variability, i.e., the P. ‘rebeli’ ecotype was less polymorphic, and its populations were much more differentiated (FST: 0.632 for EF1‐α and 0.504 for microsatellites) than the P. ‘alcon’ ecotype (0.177 and 0.082, respectively). 5. Our microsatellite data suggest that all populations of P. ‘alcon’ form a single clade but that P ‘rebeli’ can be split into either six or two clades. The former model would indicate many independent origins, especially in the mountainous areas of southern Poland. The latter, not mutually exclusive, grouping clearly reflects the use of different host ants.  相似文献   
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Secretory proteins perform a variety of important “remote-control” functions for bacterial survival in the environment. The availability of complete genome sequences has allowed us to make predictions about the composition of bacterial machinery for protein secretion as well as the extracellular complement of bacterial proteomes. Recently, the power of proteomics was successfully employed to evaluate genome-based models of these so-called secretomes. Progress in this field is well illustrated by the proteomic analysis of protein secretion by the gram-positive bacterium Bacillus subtilis, for which ~90 extracellular proteins were identified. Analysis of these proteins disclosed various “secrets of the secretome,” such as the residence of cytoplasmic and predicted cell envelope proteins in the extracellular proteome. This showed that genome-based predictions reflect only ~50% of the actual composition of the extracellular proteome of B. subtilis. Importantly, proteomics allowed the first verification of the impact of individual secretion machinery components on the total flow of proteins from the cytoplasm to the extracellular environment. In conclusion, proteomics has yielded a variety of novel leads for the analysis of protein traffic in B. subtilis and other gram-positive bacteria. Ultimately, such leads will serve to increase our understanding of virulence factor biogenesis in gram-positive pathogens, which is likely to be of high medical relevance.  相似文献   
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