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141.
Sandra Wydau Guillaume van der Rest Caroline Aubard Pierre Plateau Sylvain Blanquet 《The Journal of biological chemistry》2009,284(21):14096-14104
Several l-aminoacyl-tRNA synthetases can transfer a
d-amino acid onto their cognate tRNA(s). This harmful reaction is
counteracted by the enzyme d-aminoacyl-tRNA deacylase. Two distinct
deacylases were already identified in bacteria (DTD1) and in archaea (DTD2),
respectively. Evidence was given that DTD1 homologs also exist in nearly all
eukaryotes, whereas DTD2 homologs occur in plants. On the other hand, several
bacteria, including most cyanobacteria, lack genes encoding a DTD1 homolog.
Here we show that Synechocystis sp. PCC6803 produces a third type of
deacylase (DTD3). Inactivation of the corresponding gene (dtd3)
renders the growth of Synechocystis sp. hypersensitive to the
presence of d-tyrosine. Based on the available genomes, DTD3-like
proteins are predicted to occur in all cyanobacteria. Moreover, one or several
dtd3-like genes can be recognized in all cellular types, arguing in
favor of the nearubiquity of an enzymatic function involved in the defense of
translational systems against invasion by d-amino acids.Although they are detected in various living organisms (reviewed in Ref.
1), d-amino acids
are thought not to be incorporated into proteins, because of the
stereospecificity of aminoacyl-tRNA synthetases and of the translational
machinery, including EF-Tu and the ribosome
(2). However, the
discrimination between l- and d-amino acids by
aminoacyl-tRNA synthetases is not equal to 100%. Significant
d-aminoacylation of their cognate tRNAs by Escherichia
coli tyrosyl-, tryptophanyl-, aspartyl-, lysyl-, and histidyl-tRNA
synthetases has been characterized in vitro
(3–9).
Recently, using a bacterium, transfer of d-tyrosine onto
tRNATyr was shown to occur in vivo
(10).With such misacylation reactions, the resulting
d-aminoacyl-tRNAs form a pool of metabolically inactive molecules,
at best. At worst, d-aminoacylated tRNAs infiltrate the protein
synthesis machinery. Although the latter harmful possibility has not yet been
firmly established, several cells were shown to possess a
d-tyrosyl-tRNA deacylase, or DTD, that should help them counteract
the accumulation of d-aminoacyl-tRNAs. This enzyme shows a broad
specificity, being able to remove various d-aminoacyl moieties from
the 3′-end of a tRNA
(4–6,
11). Such a function makes the
deacylase a member of the family of enzymes capable of editing in
trans mis-aminoacylated tRNAs. This family includes several homologs
of aminoacyl-tRNA synthetase editing domains
(12), as well as peptidyl-tRNA
hydrolase (13,
14).Two distinct deacylases have already been discovered. The first one, called
DTD1, is predicted to occur in most bacteria and eukaryotes (see
d-amino acids, including
d-tyrosine (6). In
fact, in an E. coli Δdtd strain grown in the presence
of 2.4 mm d-tyrosine, as much as 40% of the cellular
tRNATyr pool becomes esterified with d-tyrosine
(10).
Open in a separate windowHomologs of dtd/DTD1 are not found in the available archaeal
genomes except that of Methanosphaera stadtmanae. A search for
deacylase activity in Sulfolobus solfataricus and Pyrococcus
abyssi led to the detection of another enzyme (DTD2), completely
different from the DTD1 protein
(15). Importing dtd2
into E. coli functionally compensates for dtd deprivation.
As shown in 16).Several cells contain neither dtd nor dtd2 homologs
(d-tyrosyl-tRNA deacylase
(DTD3). This protein, encoded by dtd3, behaves as a metalloenzyme.
Sensitivity of the growth of Synechocystis to external
d-tyrosine is strongly exacerbated by the disruption of
dtd3. Moreover, expression of the Synechocystis DTD3 in a
Δdtd E. coli strain, from a plasmid, restores the resistance of
the bacterium to d-tyrosine. Finally, using the available genomes,
we examined the occurrence of DTD3 in the living world. The prevalence of
DTD3-like proteins is surprisingly high. It suggests that the defense of
protein synthesis against d-amino acids is universal. 相似文献
TABLE 1
Distribution of DTD1 and DTD2 homologs in various phylogenetic groupsHomologs of DTD1 and DTD2 were searched for using a genomic Blast analysis against complete genomes in the NCBI Database (www.ncbi.nlm.nih.gov). Values in the table are number of species. For instance, E. coli is counted only once in γ-proteobacteria despite the fact that several E. coli strains have been sequenced.DTD1 | DTD2 | DTD1 + DTD2 | None | |
---|---|---|---|---|
Bacteria | ||||
Acidobacteria | 2 | 0 | 0 | 0 |
Actinobacteria | 27 | 0 | 0 | 8 |
Aquificae | 1 | 0 | 0 | 0 |
Bacteroidetes/Chlorobi | 12 | 0 | 0 | 5 |
Chlamydiae | 1 | 0 | 0 | 6 |
Chloroflexi | 4 | 0 | 0 | 0 |
Cyanobacteria | 5 | 0 | 0 | 16 |
Deinococcus/Thermus | 4 | 0 | 0 | 0 |
Firmicutes | ||||
Bacillales | 19 | 0 | 0 | 0 |
Clostridia | 19 | 0 | 0 | 0 |
Lactobacillales | 23 | 0 | 0 | 0 |
Mollicutes | 0 | 0 | 0 | 15 |
Fusobacteria/Planctomycetes | 2 | 0 | 0 | 0 |
Proteobacteria | ||||
α | 6 | 0 | 0 | 55 |
β | 24 | 0 | 0 | 11 |
γ | 80 | 0 | 0 | 8 |
δ | 15 | 0 | 0 | 0 |
ε | 1 | 0 | 0 | 12 |
Spirochaetes | 0 | 0 | 0 | 7 |
Thermotogae | 5 | 0 | 0 | 0 |
Archaea | ||||
Crenarchaeota | 0 | 13 | 0 | 0 |
Euryarchaeota | 1 | 26 | 0 | 2 |
Nanoarchaeota | 0 | 0 | 0 | 1 |
Eukaryota | ||||
Dictyosteliida | 1 | 0 | 0 | 0 |
Fungi/Metazoa | ||||
Fungi | 13 | 0 | 0 | 1 |
Metazoa | 19 | 0 | 0 | 0 |
Kinetoplastida | 3 | 0 | 0 | 0 |
Viridiplantae | 4 | 4 | 4 | 0 |
142.
143.
The structure of type IX collagen 总被引:26,自引:0,他引:26
M van der Rest R Mayne Y Ninomiya N G Seidah M Chretien B R Olsen 《The Journal of biological chemistry》1985,260(1):220-225
We present a detailed analysis both of tryptic peptides and amino-terminal sequences of the subunits of two collagenous fragments (HMW and LMW) previously isolated from pepsin extracts of chicken cartilage (Reese, C.A., and Mayne, R. (1981) Biochemistry 20, 5443-5448). This analysis and a comparison with the nucleotide sequence of the cDNApYN1738 (Ninomiya, Y., and Olsen, B.R. (1984) Proc. Natl. Acad. Sci. U. S. A. 81, 3014-3018) shows that HMW and LMW are pepsin-resistant fragments of a unique collagen composed of molecules with three different polypeptide chains (alpha-chains). This collagen has been assigned the type number IX, and the alpha-chain encoded by pYN1738 has been given the designation alpha 1 (IX). Type IX collagen contains three triple-helical domains and at least two sets of interchain disulfide bridges. At the amino and carboxyl ends are noncollagenous domains which do not appear to be homologous to amino and carboxyl propeptides of interstitial collagens. 相似文献
144.
The developmentally regulated type X collagen gene contains a long open reading frame without introns 总被引:15,自引:0,他引:15
Y Ninomiya M Gordon M van der Rest T Schmid T Linsenmayer B R Olsen 《The Journal of biological chemistry》1986,261(11):5041-5050
Type X collagen is a recently discovered product of hypertrophic chondrocytes that is localized to presumptive mineralization zones of hyaline cartilage. Thus, in the epiphyseal growth plate of long bones it is present only in the zone of hypertrophic chondrocytes and absent in the resting and rapidly growing cartilage and in bone. Type X collagen represents, therefore, a transient and developmentally regulated collagen which is synthesized by a subpopulation of chondrocytes. We report here the isolation and characterization of cDNA and genomic clones specific for the chicken protein. The results demonstrate that the polypeptide chains of this collagen contain three distinct domains: a short non-collagenous, amino-terminal region, a collagenous domain of 460 amino acid residues, and a non-collagenous, carboxyl-terminal domain of 170 amino acid residues. The nucleotide sequence of the gene shows that these domains are encoded by a long open reading frame that is not interrupted by introns. Examination of the amino acid sequence derived from this nucleotide sequence reveals the presence of a hydrophobic segment localized 10 amino acid residues upstream from the translational stop codon. The length and sequence characteristics of this segment raise the intriguing possibility that Type X collagen polypeptides may contain a transmembrane segment. 相似文献
145.
Identification of the type IX collagen polypeptide chains. The alpha 2(IX) polypeptide carries the chondroitin sulfate chain(s) 总被引:6,自引:0,他引:6
S Huber M van der Rest P Bruckner E Rodriguez K H Winterhalter L Vaughan 《The Journal of biological chemistry》1986,261(13):5965-5968
Type IX collagen has recently been shown to contain glycosaminoglycan chain(s) and furthermore to be immunologically identical with proteoglycan Lt (Vaughan, L., Winterhalter, K. H., and Bruckner, P. (1985) J. Biol. Chem. 260, 4758-4763). Here we demonstrate that the chondroitin sulfate carrying 115-kDa polypeptide of type IX collagen corresponds to the alpha 2(IX) chain. In addition the 84- and 68-kDa polypeptides were identified as the alpha 1(IX) and the alpha 3(IX) chains, respectively. This conclusion is based on a comparison of the tryptic fingerprints of the 84-, 115-, and 68-kDa chains of type IX collagen on high performance liquid chromatography with the similarly treated C2, C3, and C5 chains of the peptic fragment HMW. In addition, we provide evidence that both the C3 and C4 components of HMW are derived from the alpha 2(IX) chain. 相似文献
146.
Evidence from nuclear sequences that invariable sites should be considered when sequence divergence is calculated 总被引:3,自引:1,他引:2
It has long been known, from the distribution of multiple amino acid
replacements, that not all amino acids of a sequence are replaceable. More
recently, the phenomenon was observed at the nucleotide level in
mitochondrial DNA even after allowing for different rates of transition and
transversion substitutions. We have extended the search to globin gene
sequences from various organisms, with the following results: (1) Nearly
every data set showed evidence of invariable nucleotide positions. (2) In
all data sets, substitution rates of transversions and transitions were
never in the ratio of 2/1, and rarely was the ratio even constant. (3) Only
rarely (e.g., the third codon position of beta hemoglobins) was it possible
to fit the data set solely by making allowance for the number of invariable
positions and for the relative rates of transversion and transition
substitutions. (4) For one data set (the second codon position of beta
hemoglobins) we were able to simulate the observed data by making the
allowance in (3) and having the set of covariotides (concomitantly variable
nucleotides) be small in number and be turned over in a stochastic manner
with a probability that was appreciable. (5) The fit in the latter case
suggests, if the assumptions are correct and at all common, that current
procedures for estimating the total number of nucleotide substitutions in
two genes since their divergence from their common ancestor could be low by
as much as an order of magnitude. (6) The fact that only a small fraction
of the nucleotide positions differ is no guarantee that one is not
seriously underestimating the total amount of divergence (substitutions).
(7) Most data sets are so heterogeneous in their number of transition and
transversion differences that none of the current models of nucleotide
substitution seem to fit them even after (a) segregation of coding from
noncoding sequences and (b) splitting of the codon into three subsets by
codon position. (8) These frequently occurring problems cannot be seen
unless several reasonably divergent orthologous genes are examined
together.
相似文献
147.
Native type IV collagen was isolated from human placental tissue by pepsin digestion, fractional salt precipitation, reduction and alkylation, a second pepsin digestion, and chromatography on diethylaminoethyl- and carboxymethyl-cellulose. After denaturation, 10 distinct peptides were isolated from this material by molecular sieve, ion-exchange, and high-performance liquid chromatography. All of the peptides were found to have amino acid compositions characteristic of type IV collagen. Analysis of the eight major peptides by amino-terminal amino acid sequencing and by cyanogen bromide and tryptic peptide mapping has revealed the manner in which they are derived from type IV collagen. Pepsin liberates two large peptides by attacking non-triple-helical regions, one derived from the alpha 1 (IV) chain (F2, Mr 90 000) and one derived from the alpha 2 (IV) chain (F3, Mr 75 000). The alpha 1 (IV)-derived F2 peptide is also represented in the pepsin digest by amino-terminal and carboxy-terminal subfragments [F4c (Mr 41 000) and F4a (Mr 60 000)], as is the alpha 2 (IV)-derived F3 peptide [F5 (Mr 28 000) and F4b (Mr 50 000), respectively]. These findings indicate that the molecular regions from which the larger peptides are derived in themselves contain pepsin-sensitive (non-triple-helical) domains. In addition, several of the peptides examined were found to be present in two slightly different forms, suggesting that closely adjacent pepsin-sensitive sites often exist within the type IV collagen molecules. The methods outlined here provide a reliable means by which identifiable type IV collagen peptides can be isolated.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
148.
Twenty- two pigs with an average weight of 24.2 kg were divided into two groups. Five grams of an ointment containing 40% of the n-3 polyunsaturated fatty acids, eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3) as ethylesters, was administered cutaneously to the experimental group and orally to the control group twice daily over a 3 day period. Surgical biopsies of subcutaneous fat of all 22 pigs were taken before and after the ethylester administration period. Blood samples were taken from the jugular vein. Skin biopsies were taken after the application period, and the presence of pathological features and inter-animal variation were noted. No increase of EPA and DHA in subcutaneous fat was found in either group. In the oral group the content of EPA had increased in the total plasma lipids on the first day after the end of the experiment. It is concluded that a rapid transcutaneous absorption of EPA and DHA as ethylesters does not seem to occur in pigs. 相似文献
149.
Secondary transporters for citrate and the Mg(2+)-citrate complex in Bacillus subtilis are homologous proteins.
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Citrate uptake in Bacillus subtilis is mediated by a secondary transporter that transports the complex of citrate and divalent metal ions. The gene coding for the transporter termed CitM was cloned, sequenced, and functionally expressed in Escherichia coli. Translation of the base sequence to the primary sequence revealed a transporter that is not homologous to any known secondary transporter. However, CitM shares 60% sequence identity with the gene product of open reading frame N15CR that is on the genome of B. subtilis and for which no function is known. The hydropathy profiles of the primary sequences of CitM and the unknown gene product are very similar, and secondary structure prediction algorithms predict 12 transmembrane-spanning segments for both proteins. Open reading frame N15CR was cloned and expressed in E. coli and was shown to be a citrate transporter as well. The transporter is termed CitH. A remarkable difference between the two transporters is that citrate uptake by CitM is stimulated by the presence of Mg2+ ions, while citrate uptake by CitH is inhibited by Mg2+. It is concluded that the substrate of CitM is the Mg(2+)-citrate complex and that CitH transports the free citrate anion. Uptake experiments in right-side-out membrane vesicles derived from E. coli cells expressing either CitM or CitH showed that both transporters catalyze electrogenic proton/substrate symport. 相似文献
150.
E Aubert-Foucher B Font D Eichenberger D Goldschmidt C Lethias M van der Rest 《The Journal of biological chemistry》1992,267(22):15759-15764
A new molecule, type XIV collagen, with domains homologous to type IX and XII collagens has been recently discovered in pepsin extracts of fetal bovine tissues (Dublet, B., and van der Rest, M. (1991) J. Biol. Chem. 266, 6853-6858). In the present study, we describe the purification and the characterization of the intact native form of this newly discovered collagen. By using only two chromatographic steps we were able to obtain pure type XIV collagen. Furthermore, minor modifications of the protocol allowed us to perform the simultaneous large scale purification of type XII and type XIV collagens from the same tissue. Intact type XIV collagen migrates on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as two bands of 220 and 290 kDa (reducing conditions). After collagenase treatment, a single band of 190 kDa is observed, which represents the large non-collagenous domain of the molecule (NC3). Rotary shadowing electron micrographs of intact type XIV collagen show a cross-shaped structure formed by a thin tail attached through a central globule to three identical "fingers." These properties are similar to those previously described for intact chicken type XII collagen (Dublet, B., Oh, S., Sugrue, S. P., Gordon, M. K., Gerecke, D. R., Olsen, B. R., and van der Rest, M. (1989) J. Biol. Chem. 264, 13150-13156), but the two molecules are different gene products and have charge and glycosylation differences. Finally, we show that the three chains of purified type XIV collagen have an apparent molecular mass of approximately 220 kDa and are not cross-linked to each other by bonds other than disulfide bridges. The same observation was made for type XII collagen. In both cases, the 290-kDa migrating band in SDS-PAGE is due to incomplete denaturation in electrophoresis sample buffer in the absence of urea. 相似文献