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Replication slippage may cause parallel evolution in the secondary structures of mitochondrial transfer RNAs 总被引:9,自引:4,他引:5
Presence of the dihydrouridine (D) stem in the mitochondrial cysteine tRNA
is unusually variable among lepidosaurian reptiles. Phylogenetic and
comparative analyses of cysteine tRNA gene sequences identify eight
parallel losses of the D-stem, resulting in D-arm replacement loops.
Sampling within the monophyletic Acrodonta provides no evidence for
reversal. Slipped-strand mispairing of noncontiguous repeated sequences
during replication or direct replication slippage can explain repeats
observed within cysteine tRNAs that contain a D-arm replacement loop. These
two mechanisms involving replication slippage can account for the loss of
the cysteine tRNA D-stem in several lepidosaurian lineages, and may
represent general mechanisms by which the secondary structures of
mitochondrial tRNAs are altered.
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4.
The complete cDNA sequence and protein reading frame of a developmentally
regulated hemocyanin subunit in the Dungeness crab (Cancer magister) is
presented. The protein sequence is aligned with 18 potentially homologous
hemocyanin-type proteins displaying apparent sequence similarities.
Functional domains are identified, and a comparison of predicted
hydrophilicities, surface probabilities, and regional backbone
flexibilities provides evidence for a remarkable degree of structural
conservation among the proteins surveyed. Parsimony analysis of the protein
sequence alignment identifies four monophyletic groups on the arthropodan
branch of the hemocyanin gene tree: crustacean hemocyanins, insect
hexamerins, chelicerate hemocyanins, and arthropodan prophenoloxidases.
They form a monophyletic group relative to molluscan hemocyanins and
nonarthropodan tyrosinases. Arthropodan prophenoloxidases, although
functionally similar to tyrosinases, appear to belong to the arthropodan
hexamer- type hemolymph proteins as opposed to molluscan hemocyanins and
tyrosinases.
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5.
Proliferating cell nuclear antigen (PCNA) is an essential protein for DNA replication, DNA repair, cell cycle regulation, chromatin remodeling, and epigenetics. Many proteins interact with PCNA through the PCNA interacting peptide (PIP)-box or the newly identified AlkB homolog 2 PCNA interacting motif (APIM). The xeroderma pigmentosum group A (XPA) protein, with a central but somewhat elusive role in nucleotide excision repair (NER), contains the APIM sequence suggesting an interaction with PCNA. With an in vivo based approach, using modern techniques in live human cells, we show that APIM in XPA is a functional PCNA interacting motif and that efficient NER of UV lesions is dependent on an intact APIM sequence in XPA. We show that XPA−/− cells complemented with XPA containing a mutated APIM sequence have increased UV sensitivity, reduced repair of cyclobutane pyrimidine dimers and (6–4) photoproducts, and are consequently more arrested in S phase as compared to XPA−/− cells complemented with wild type XPA. Notably, XPA colocalizes with PCNA in replication foci and is loaded on newly synthesized DNA in undamaged cells. In addition, the TFIIH subunit XPD, as well as XPF are loaded on DNA together with XPA, and XPC and XPG colocalize with PCNA in replication foci. Altogether, our results suggest a presence of the NER complex in the vicinity of the replisome and a novel role of NER in post-replicative repair. 相似文献
6.
Hagen L Kavli B Sousa MM Torseth K Liabakk NB Sundheim O Pena-Diaz J Otterlei M Hørning O Jensen ON Krokan HE Slupphaug G 《The EMBO journal》2008,27(1):51-61
Human UNG2 is a multifunctional glycosylase that removes uracil near replication forks and in non-replicating DNA, and is important for affinity maturation of antibodies in B cells. How these diverse functions are regulated remains obscure. Here, we report three new phosphoforms of the non-catalytic domain that confer distinct functional properties to UNG2. These are apparently generated by cyclin-dependent kinases through stepwise phosphorylation of S23, T60 and S64 in the cell cycle. Phosphorylation of S23 in late G1/early S confers increased association with replication protein A (RPA) and replicating chromatin and markedly increases the catalytic turnover of UNG2. Conversely, progressive phosphorylation of T60 and S64 throughout S phase mediates reduced binding to RPA and flag UNG2 for breakdown in G2 by forming a cyclin E/c-myc-like phosphodegron. The enhanced catalytic turnover of UNG2 p-S23 likely optimises the protein to excise uracil along with rapidly moving replication forks. Our findings may aid further studies of how UNG2 initiates mutagenic rather than repair processing of activation-induced deaminase-generated uracil at Ig loci in B cells. 相似文献
7.
Human AlkB homolog 1 is a mitochondrial protein that demethylates 3-methylcytosine in DNA and RNA 总被引:1,自引:0,他引:1
Westbye MP Feyzi E Aas PA Vågbø CB Talstad VA Kavli B Hagen L Sundheim O Akbari M Liabakk NB Slupphaug G Otterlei M Krokan HE 《The Journal of biological chemistry》2008,283(36):25046-25056
The Escherichia coli AlkB protein and human homologs hABH2 and hABH3 are 2-oxoglutarate (2OG)/Fe(II)-dependent DNA/RNA demethylases that repair 1-methyladenine and 3-methylcytosine residues. Surprisingly, hABH1, which displays the strongest homology to AlkB, failed to show repair activity in two independent studies. Here, we show that hABH1 is a mitochondrial protein, as demonstrated using fluorescent fusion protein expression, immunocytochemistry, and Western blot analysis. A fraction is apparently nuclear and this fraction increases strongly if the fluorescent tag is placed at the N-terminal end of the protein, thus interfering with mitochondrial targeting. Molecular modeling of hABH1 based upon the sequence and known structures of AlkB and hABH3 suggested an active site almost identical to these enzymes. hABH1 decarboxylates 2OG in the absence of a prime substrate, and the activity is stimulated by methylated nucleotides. Employing three different methods we demonstrate that hABH1 demethylates 3-methylcytosine in single-stranded DNA and RNA in vitro. Site-specific mutagenesis confirmed that the putative Fe(II) and 2OG binding residues are essential for activity. In conclusion, hABH1 is a functional mitochondrial AlkB homolog that repairs 3-methylcytosine in single-stranded DNA and RNA. 相似文献
8.
Microsatellite allele frequencies in humans and chimpanzees, with implications for constraints on allele size 总被引:24,自引:6,他引:18
The distributions of allele sizes at eight simple-sequence repeat (SSR) or
microsatellite loci in chimpanzees are found and compared with the
distributions previously obtained from several human populations. At
several loci, the differences in average allele size between chimpanzees
and humans are sufficiently small that there might be a constraint on the
evolution of average allele size. Furthermore, a model that allows for a
bias in the mutation process shows that for some loci a weak bias can
account for the observations. Several alleles at one of the loci (Mfd 59)
were sequenced. Differences between alleles of different lengths were found
to be more complex than previously assumed. An 8-base-pair deletion was
present in the nonvariable region of the chimpanzee locus. This locus
contains a previously unrecognized repeated region, which is imperfect in
humans and perfect in chimpanzees. The apparently greater opportunity for
mutation conferred by the two perfect repeat regions in chimpanzees is
reflected in the higher variance in repeat number at Mfd 59 in chimpanzees
than in humans. These data indicate that interspecific differences in
allele length are not always attributable to simple changes in the number
of repeats.
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9.
Monoclonal antibodies against chicken type V collagen: production, specificity, and use for immunocytochemical localization in embryonic cornea and other organs 总被引:23,自引:17,他引:6 下载免费PDF全文
TF Linsenmayer JM Fitch TM Schmid Zak NB E Gibney RD Sanderson R Mayne 《The Journal of cell biology》1983,96(1):124-132
Two monoclonal antibodies have been produced against chick type V collagen and shown to be highly specific for separate, conformational dependent determinants within this molecule. When used for immunocytochemical tissue localization, these antibodies show that a major site for the in situ deposition of type V is within the extracellular matrices of many dense connective tissues. In these, however, it is largely in a form unavailable to the antibodies, thus requiring a specific “unmasking” treatment to obtain successful immunocytochemical staining. The specificity of these two IgG antibodies was determined by inhibition ELISA, in which only type V and no other known collagen shows inhibition. In ELISA, mixtures of the two antibodies give an additive binding reaction to the collagen, suggesting that each is against a different antigenic determinant. That both antigenic determinants are conformational dependent, being either in, or closely associated with, the collagen helix is demonstrated by the loss of antibody binding to molecules that have been thermally denatured. The temperature at which this occurs, as assayed by inhibition ELISA, is very similar to that at which the collagen helix melts, as determined by optical rotation. This gives strong additional evidence that the antibodies are directed against the collagen. The antibodies were used for indirect immunofluorescence analyses of cryostat sections of corneas and other organs from 17 to 18-day-old chick embryos. Of all tissues examined only Bowman’s membrane gave a strong staining reaction with cryostat sections of unfixed material. Staining in other areas of the cornea and in other tissues was very light or nonexistent. When, however, sections were pretreated with pepsin dissolved in dilute HAc or, surprisingly, with the dilute HAc itself dramatic new staining by the antibodies was observed in most tissues examined. The staining, which was specific for the anti-type V collagen antibodies, was largely confined to extracellular matrices of dense connective tissues. Experiments using protease inhibitors suggested that the “unmasking” did not involve proteolysis. We do not yet know the mechanism of this unmasking; however, one possibility is that the dilute acid causes swelling or conformational changes in a type-V collagen-containing supramolecular structure. Further studies should allow us to determine whether this is the case. 相似文献
10.
Hunter NB Moseley Andrew N Lane Alex C Belshoff Richard M Higashi Teresa WM Fan 《BMC biology》2012,10(1):1-2
This article is a response to Wang and Luo. See correspondence article http://www.biomedcentral.com/1741-7007/10/30/ [WEBCITE] and the original research article http://www.biomedcentral.com/1741-7007/9/24 [WEBCITE]. 相似文献