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131.
In vitro replication assays for detection and quantification of bypass of UV-induced DNA photoproducts were used to compare the capacity of extracts prepared from different human cell lines to replicate past the cis,syn cyclobutane thymine dimer ([c,s]TT). The results demonstrated that neither nucleotide excision repair (NER) nor mismatch repair (MMR) activities in the intact cells interfered with measurements of bypass replication efficiencies in vitro. Extracts prepared from HeLa (NER- and MMR-proficient), xeroderma pigmentosum group A (NER-deficient), and HCT116 (MMR-deficient) cells displayed similar capacity for translesion synthesis, when the substrate carried the site-specific [c,s]TT on the template for the leading or the lagging strand of nascent DNA. Extracts from xeroderma pigmentosum variant cells, which lack DNA polymerase eta, were devoid of bypass activity. Bypass-proficient extracts as a group (n=16 for 3 extracts) displayed higher efficiency (P=0.005) for replication past the [c,s]TT during leading strand synthesis (84+/-22%) than during lagging strand synthesis (64+/-13%). These findings are compared to previous results concerning the bypass of the (6-4) photoproduct [Biochemistry 40 (2001) 15215] and analyzed in the context of the reported characteristics of bypass DNA polymerases implicated in translesion synthesis of UV-induced DNA lesions. Models to explain how these enzymes might interact with the DNA replication machinery are considered. An alternative pathway of bypass replication, which avoids translesion synthesis, and the mutagenic potential of post-replication repair mechanisms that contribute to the duplication of the human genome damaged by UV are discussed. 相似文献
132.
Platelet-derived growth factor exerts disparate effects on odontoblast differentiation depending on the dimers in rat dental pulp cells 总被引:3,自引:0,他引:3
Yokose S Kadokura H Tajima N Hasegawa A Sakagami H Fujieda K Katayama T 《Cell and tissue research》2004,315(3):375-384
Platelet-derived growth factor (PDGF) has recently been demonstrated to control the expression of alkaline phosphatase and proteoglycan synthesis of odontoblastic cells in dental pulp tissues. Although PDGF appears to be closely related to dentinogenesis, much about the mode of action of PDGF on odontoblast differentiation remains unclear. In this study, we examined the effects of three PDGF dimers (PDGF AA, AB, and BB) on odontoblastic differentiation of dental pulp cells in long-term mineralized cultures. Dental pulp cells isolated from rat lower incisors were continuously treated with each of PDGF AA, AB, and BB in separate cultures for 20 days. The three PDGF dimers suppressed alkaline phosphatase activity, osteocalcin and calcium content, and the formation of dentin-like nodules. The expression of mRNA for dentin sialoprotein (DSP) in the cells was inhibited by PDGF AA treatment, whereas PDGF AB and BB treatment stimulated the expression of DSP, even though the dentin-like nodule formation was inhibited. Although the effects of PDGF on odontoblastic differentiation varied among the dimers, the cells expressed both PDGF and receptors, whose quantities were similar. These results suggest that PDGF exerts diverse effects on odontoblastic differentiation depending on its dimeric form. These in vitro findings explain, at least in part, the in vivo action of PDGF in dentinogenesis during the repair process of damaged dental pulp.This work was supported in part by grants-in-aid from the Ministry of Science, Education, and Culture of Japan 相似文献
133.
Dansette PM Bertho G Mansuy D 《Biochemical and biophysical research communications》2005,338(1):450-455
Oxidation of 2-phenylthiophene (2PT) by rat liver microsomes, in the presence of NADPH and glutathione (GSH), led to three kinds of metabolites whose structures were established by 1H NMR and mass spectrometry. The first ones were 2PT-S-oxide dimers formed by Diels-Alder type dimerization of 2PT-S-oxide, while the second ones were GSH adducts derived from the 1,4-Micha?l-type addition of GSH to 2PT-S-oxide. The third metabolites were GSH adducts resulting from a nucleophilic attack of GSH to the 4,5-epoxide of 2PT. Oxidation of 2PT by recombinant, human cytochrome P4501A1, in the presence of NADPH and GSH, also led to these three kinds of metabolites. These results provide the first evidence that cytochrome P450 may catalyze the oxidation of thiophene compounds with the simultaneous formation of two reactive intermediates, a thiophene-S-oxide and a thiophene epoxide. 相似文献
134.
Inoki T Yamagami S Inoki Y Tsuru T Hamamoto T Kagawa Y Mori T Endo H 《Biochemical and biophysical research communications》2004,314(4):1036-1043
Damaged DNA-binding protein (DDB) is a heterodimer (DDB1 and DDB2), which is implicated in the repair of UV-irradiated DNA damage. Here we have identified four DDB2 variants from HeLa cells (D1-D4) that are generated by alternative splicing. Analysis of tissue distribution by RT-PCR indicates that D1 is the most highly expressed in human brain and heart. A DNA repair assay revealed that both D1 and D2 are dominant negative inhibitors. Electrophoresis mobility shift assays indicated that D1 and D2 are not part of the damaged DNA-protein complex. Co-immunoprecipitation studies show that DDB2-WT interacts with D1 and itself. Nuclear import of DDB1 was less induced by transfection with D1 than WT. Based on these results, D1 and D2 are dominant negative inhibitors of DNA repair, which is probably due to disruption of complex formation between DDB1 and DDB2-WT and of DDB1 nuclear import. 相似文献
135.
Reyes-Vivas H Martínez-Martínez E Mendoza-Hernández G López-Velázquez G Pérez-Montfort R Tuena de Gómez-Puyou M Gómez-Puyou A 《Proteins》2002,48(3):580-590
The susceptibility to subtilisin of homodimeric triosephosphate isomerase from Trypanosoma brucei (TbTIM) and Trypanosoma cruzi (TcTIM) was studied. Their amino sequence and 3D structure are markedly similar. In 36 h of incubation at a molar ratio of 4 TIM per subtilisin, TcTIM underwent extensive hydrolysis, loss of activity, and large structural alterations. Under the same conditions, only about 50% of the monomers of TbTIM were cleaved in two sites. The higher sensitivity of TcTIM to subtilisin is probably due to a higher intrinsic flexibility. We isolated and characterized TbTIM that had been exposed to subtilisin. It exhibited the molecular mass of the dimer, albeit it was formed by one intact and one nicked monomer. Its k(cat) with glyceraldehyde 3-phosphate was half that of native TbTIM, with no change in K(m). The intrinsic fluorescence of nicked TbTIM was red-shifted by 5 nm. The association between subunits was not affected. The TbTIM data suggest that there are structural differences in the two monomers or that alterations of one subunit change the characteristics of the other subunit. In comparison to the action of subtilisin on TIMs from other species, the trypanosomal enzymes appear to be unique. 相似文献
136.
137.
Michael A. Resnick James Westmoreland Enrique Amaya Kerry Bloom 《Molecular & general genetics : MGG》1987,210(1):16-22
Summary The centromere is the region within a chromosome that is required for proper segregation during mitosis and meiosis. Lesions in this sequence represent a unique type of damage, as loss of function could result in catastrophic loss of the genetic material of an entire chromosome. We have measured the induction by ultraviolet (UV) light of pyrimidine dimers in a 2550-bp restriction fragment that includes the centromere region of chromosome III in Saccharomyces cerevisiae. Yeast cells were exposed to ultraviolet light, cellular DNA was gently extracted, and subsequently treated with a UV-specific endonuclease to cleave all pyrimidine dimers. The sites of UV-specific nuclease scission within the centromere were determined by separating the DNA according to molecular weight, transferring the fragments to nitrocellulose, and hybridizing to a radiolabeled 624-bp fragment homologous to the centromere DNA from chromosome III. Several hotspots were identified in chromatin DNA from cells, as well as in irradiated deproteinized DNA. Double strand damage due to closely opposed pyrimidine dimers was also observed. At biological doses (35% survival) there are approximately 0.1 to 0.2 pyrimidine dimers per centromere. These dimers are efficiently repaired in the centromere and surrounding region. 相似文献
138.
A. A. Krasnovsky Jr. J. Lopez P. Cheng P. A. Liddell R. E. Blankenship T. A. Moore D. Gust 《Photosynthesis research》1994,40(2):191-198
Both photogeneration and quenching of singlet oxygen by monomeric and aggregated (dimeric and oligomeric) molecules of bacteriochlorophyll (BChl) d have been studied in solution and in chlorosomes isolated from the green photosynthetic bacterium Chlorobium vibrioforme f. thiosulfatophilum. The yield of singlet-oxygen photogeneration by pigment dimers was about 6 times less than for monomers. Singlet oxygen formation was not observed in oligomer-containing solutions or in chlorosomes. To estimate the efficiency of singlet oxygen quenching an effective rate constant for 1O2 quenching by BChl molecules (kq
M) was determined using the Stern-Volmer equation and the total concentration of BChl d in the samples. In solutions containing only monomeric BChl, the kq
M values coincide with the real values for 1O2 quenching rate constants by BChl molecules. Aggregation weakly influenced the kq
M values in pigment solutions. In chlorosomes (which contain both BChl and carotenoids) the kq
M value was less than in solutions of BChl alone and much less than in acetone extracts from chlorosomes. Thus 1O2 quenching by BChl and carotenoids is much less efficient in chlorosomes than in solution and is likely caused primarily by BChl molecules which are close to the surface of the large chlorosome particles. The data allow a general conclusion that monomeric and dimeric chlorophyll molecules are the most likely sources of 1O2 formation in photosynthetic systems and excitation energy trapping by the long wavelength aggregates as well as 1O2 physical quenching by monomeric and aggregated chlorophyll can be considered as parts of the protective system against singlet oxygen formation.Abbreviations BChl
bacteriochlorophyll
- MBpd
methyl bacteriopheophorbide
- Chl
chlorophyll
- TPP
meso-tetraphenylporphyrin
- TPPS
meso-tetra (p-sulfophenyl) porphyrin 相似文献
139.
Jane S. Murray M. Edward Grice Peter Politzer Margaret C. Etter 《Molecular Engineering》1991,1(1):75-87
We have carried out anab initio self-consistent-field molecular orbital analysis of the structures, atomic charges and electrostatic potentials of 1,3-bisphenylurea, 1,3-bis(m-nitrophenyl)urea and 1,3-bis(p-nitrophenyl)urea. Our results provide insight into the contrasting hydrogen bond patterns and preferences of these molecules in crystalline environments. The tendency for 1,3-bisphenylurea to form homomeric rather than heteromeric (i.e. host-guest) crystals is attributed to a relatively strong and extended homomeric attractive electrostatic interaction between two like molecules. Our calculated internuclear distances and atomic charges indicate that these molecules have some degree of H...O intramolecular hydrogen bonding between the carbonyl oxygen and the nearest aromatic hydrogens when the aromatic and the urea portions of the molecules are coplanar. This interaction is strongest for the meta nitro derivative, consistent with the latter remaining very nearly planar in its cocrystal structures. Our surface electrostatic potentials for the three diaryl ureas are used to interpret their differing crystallization properties and tendencies to form cocrystals with guest molecules. 相似文献
140.
Christopher W. Lawrence Swapan K. Banerjee Angela Borden J. Eugene LeClere 《Molecular & general genetics : MGG》1990,222(1):166-168
Summary The lesions produced by SOS-dependent mutagens in Escherichia coli are commonly referred to as non-pairing or non-instructive. Although these terms are likely to be appropriate for some lesions, particularly the abasic site, for others, such as the cyclobutane dimer, their suitability is open to question. To address this question, we have compared the error frequencies and spectra that result when a uniquely located T-T sequence, carried in a single-stranded vector, contains either a cis-syn or a trans-syn cyclobutane dimer, or when either the 5T or 3T is converted to an abasic site. The data suggest that the high accuracy with which the dimer-containing templates are replicated is unlikely to be the consequence of polymerase preference for the non-instructive insertion of dAMP. Similarly, mis-pairing, rather than non-pairing, is likely to cause mutations. Cyclobutane dimers seem therefore to be misinstructive rather than non-instructive lesions, and the common feature shared by SOS-inducing lesions is more their ability to block replication than inability to form correct base pairs. 相似文献