Kim, Do-Hyung;Kim, Jeong-Hoon;Park, Byoung Chul;Lee, Do Hee;Cho, Sayeon;Park, Sung Goo
Bulletin of the Korean Chemical Society
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v.35
no.10
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pp.3005-3008
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2014
Human ChlR1 protein (hChlR1), a member of the cohesion establishment factor family, plays an important role in the segregation of sister chromatids for maintenance of genome integrity. We previously reported that hChlR1 interacts with hFen1 and stimulates its nuclease activity on the flap-structured DNA substrate covered with RPA. To elucidate the relationship between hChlR1 and Okazaki fragment processing, the effect of hChlR1 on in vitro nuclease activities of hFen1 and hDna2 was examined. Independent of ATPase activity, hChlR1 stimulated endonuclease activity of hFen1 but not that of hDna2. Our findings suggest that the acceleration of Okazaki fragment processing near cohesions may aid in reducing the size of the replication machinery, thereby facilitating its entry through the cohesin ring.
Chlorophyll synthase (ChlG) and bacteriochlorophyll synthase (BchG) have a high degree of substrate specificity. The BchG mutant of Rhodobacter sphaeroides, BG1 strain, is photosynthetically incompetent. When BG1 harboring chlG of Synechocystis sp. PCC 6803 was cultured photoheterotrophically, colonies arose at a frequency of approximately 10-8. All the suppressor mutants were determined to have the same mutational change, ChlGI44F. The mutated enzyme ChlGI44F showed BchG activity. Remarkably, BchGF28I, which has the substitution of F at the corresponding 28th residue to I, showed ChlG activity. The Km values of ChlGI44F and BchGF28I for their original substrates, chlorophyllide (Chlide) a and bacteriochlorophyllide (Bchlide) a, respectively, were not affected by the mutations, but the Km values of ChlGI44F and BchGF28I for the new substrates Bchlide a and Chlide a, respectively, were more than 10-fold larger than those for their original substrates, suggesting the lower affinities for new substrates. Taken together, I44 and F28 are important for the substrate specificities of ChlG and BchG, respectively. The BchG activity of ChlGI44F and the ChlG activity of BchGF28I further suggest that ChlG and BchG are evolutionarily related enzymes.
The inhibition characteristics of chlorsulfuron [CHL, 2-chloro-N-[{ (4-methoxy-6-methyl-1,3,5-triazin-2-yl)amino}carbonyl]benzenesulfonamide] and imazaquin [IMA, 2-{4,5-dihydro-4-methyl-4-(1-methy-lethyl)-5-oxo-1H-imidazol-2-yl}-3-quinolinecarboxylic acid] on acetolactate synthase(ALS) activity of corn plants were investigated. CHL and IMA rapidly inhibited ALS activity of corn plants in vitro. Their $I_{50}$ values for ALS activity were 100nM and $5{\mu}M$, respectively, indicating that CHL had 50 times more inhibitory effect on ALS activity than IMA. The first applied herbicide had a dominant inhibitory effect on ALS activity when the two herbicides were applied sequentially. Branched-chain amino acids, valine(Val), leucine(Leu), and isoleucine(Ile) showed a feedback inhibition on ALS activity ; Val or Leu had a more inhibitory effect on ALS activity than Ile. Branchedchain amino acids and CHL or IMA exhibited an additive effect on inhibiting ALS activity. This suggests that branched-chain amino acids inhibit ALS activity by a different mechanisms) from that of CHL or IMA. Apparent ALS activity, which was measured on the basis of the conversion of pyruvate to acetolactate, was decreased by the addition of 2-ketobutyrate into the ALS reaction mixture in a concentration-dependent manner. In addition, kinetic studies revealed that CHL acts as a noncompetitive inhibitor, while IMA acts as an uncompetitive inhibitor to ALS with respect to pyruvate.
Promoters of the chlorophyll a/b bidning protein genes, cab1, and cab2, of Arabidopsis thaliana were studied for their functions in differential expression during greening of etiolated shoots. The etiolated shoots were derived from leaves of transgenic tobacco plants with the cab-CAT (chloramphenicol acetyltransferase) translational fusions, and CAT activity was measured to monitor the activities of the cab promoters. Cab1 promoter activity increased rapidly and showed saturation after about 24 hours of greening, but that of cab2 increased with about 2 day-lag period and showed saturation after 6 days. Cab1 promoter activity was more sensitive to levulinic acid (LA) compared with cab2 activity. Cab2 promoter activity was inhibited more sensitively by chloramphynicol (CAP) than by inhibitors of Chl formation. Cab1 promoter activity was, however, inhibited less sensitively by CAP than by LA. The treatment of abscisic acid (ABA) did not block Chl synthesis so significantly as LA treatment did, and cab2 promoter activity was much less sensitive to ABA compared with that of cab1. These results suggest that cab1 expression is strongly related with Chl formation, possibly with $\delta$-aminolevulinic acid accumulation, and cab2 expression is suppressed more by the blockage of translation of Chl a-apoproteins than by the blockage of Chl a accumulation.
The formation of chlorophyll-protein complexes (CP-complexes) during the greening of rape cotyledons (Brassica napus cv. Yongdang) was investigated by the SDS-polyacrylamide gel electrophoresis. The total chlorophyll content and Chl a/b ratio were also determined. In addition, the effects of dark treatment on the CP-complex patterns during greening have been examined with respect to their photosynthetic electron transport activity. Greening has brought about the increasein total chlorophyll content and the decrease in Chl a/b ratio, but there have been no changes in Chl a/b ratio after 24 hrs of greening. The light-harvesting chlorophyll a/b-protein complex (LHCP-complex0 was predominant during the initial greening period. Thereafter, the amout of chlorophyll a-protein complex (CP I-complex) was gradually increased. Twenty-four-hr dark treatment immediately after illumination for 6 hrs and 12 hrs resulted in the increase of the Chl a/b ration and the CP I complex, otherwise the decrease of the LHCP-complex. The LHCP/CP I ratio was gradually decreased with further greening, and appeared no change after 48 hrs illumination. The investigation of the photosynthetic electron transport activity indicated that photosystem (PS) II activity (H2Olongrightarrowp-PD*+FeCy**) did not change, but the activity of PS I was increased suddenly due to the dark treatment. The data suggests that the increase of CP I-complex may result in that of P-700, that is, the increase of PS I activity.
Park, Joo hyun;Choi, Jeong-Wook;Lee, Min-Kyeong;Choi, Youn Hee;Nam, Taek-Jeong
Fisheries and Aquatic Sciences
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v.22
no.1
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pp.2.1-2.9
/
2019
In the present study, the protective effects of Hizikia fusiforme and Chlorella sp. extracts on lead acetate-induced hepatotoxicity were investigated. Hepatic damage was induced in rats by intraperitoneal (i.p.) injection of lead acetate and the protective effects of H. fusiforme (HZK) and Chlorella sp. (CHL) extracts on lead acetate-induced hepatic damage in rat liver were examined. The results revealed significantly increased glutamic oxaloacetate and glutamic pyruvic transaminase levels in the group treated with lead acetate only (Pb group); oral administration of HZK and CHL extracts tended to decrease the enzyme levels similar to those observed in the control group. Regarding antioxidant enzymes, superoxide dismutase activity was increased in the Pb group and decreased in a concentration-dependent manner in the HZK- and CHL-treated groups. Glutathione levels were increased in a concentration-dependent manner in the HZK- and CHL-treated groups. There was no significant difference in catalase activity. Western blot analysis showed inflammation-related protein expression in mitogen-activated protein kinase and Nrf2 pathways was affected in the HZK- and CHL-treated groups. Therefore, HZK and CHL extracts exerted antioxidant and anti-inflammatory effects against lead acetate-induced hepatotoxicity. Development of functional health foods containing HZK and CHL extracts, which have hepatoprotective effects against inhaled lead acetate, should be considered.
The effects of cadmium ions ($Cd^{2+}$) on the Chl a fluorescence of Ricciocarpos natans were investigated in order to determine whether Chl fluorescence can be used as a biomarker to estimate the physiological responses of plants to cadmium stress. In all plants treated with $Cd^{2+}$, the image of Fv/Fm, which represents the maximum photochemical efficiency of PSII, changed as the $Cd^{2+}$ concentration increased, when treated for 48 h or more. Changes of ${\Phi}_{PSII}$ and $Q_P$ images were recognized even at 10 ${\mu}M$$Cd^{2+}$. The Chl a O-J-I-P fluorescence transient was also affected even at 10 ${\mu}M$$Cd^{2+}$. The fluorescence yield decreased considerably in steps J, I and P in plants treated with $Cd^{2+}$, although a typical polyphasic rise was observed in non-treated plants. The Chl fluorescence parameters, Fm, Fv/Fo, Sm, SFIabs, PIabs and ETo/CS, decreased as the $Cd^{2+}$ concentration increased, while the Mo and Kn parameters increased. Peroxidase activity decreased significantly and catalase activity increased as the $Cd^{2+}$ concentration increased. Because of its sensitivity to $Cd^{2+}$ Ricciocarpos natans is useful in experiments investigating the responses of plants to cadmium exposure. Several parameters (Fm, Fv/Fo, Sm, SFIabs, PIabs, ETo/CS, Mo and Kn) can be applied to determine quantitatively the physiological states of plants under cadmium stress.
Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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v.3
no.1
/
pp.11-21
/
1999
The effect of sulfite on barley seedlings was investigated through Chl content, the electron transport activity of the photosystem, and Chl fluorescence. Barley leaves were harvested every 12 hrs during greening periods, and were then treated with a sulfite solution in either light or dark conditions. In both cases, the Chl content decreased in comparison with the control at any greening period. After sulfite treatment in the light, the activity of PS I decreased slightly, yet that of PSII showed a decrease of about 15%. The values of Fv, qP and qE decreased, however, the value of ql increased compared with the control. In addition, the value of qE decreased in leaves greened more than 12 hrs compared with that of the control. This indicates that the photosynthetic complex involved in energy dependent fluorescence quenching is undeveloped in a 12 hrs greened leaf, accordingly, it was a hardly affected by sulfite. After sulfite treatment in the dark, the activities of PSII and PSI decreased slightly, there was a small change in the value of Fv, qP decreased, and qE and the ratio of qNP/q increased in comparison with the control. As a result, PSII and PSI were not inhibited, however, the redox of QA was inhibited, and the excited energy was lost through the nonphotochemical pathway. The effects of sulfite in light or dark conditions were not considerably different with the Chl fluorescence quenching analysis method. In both light and dark conditions, the value of qP significantly decreased with sulfite compared to that of the control. This implies that the redox of QA was inhibited by sulfite in both light and dark contions.
To elucidate a part of phosphorus cycle an'd the nutritional status of phosphorus. the degradation rates of organic phosphate by phosphatase activity (PA). were studied at the water column of Lake Soyang. from March 1900 to April 1991. Phosphatase activity showed the range of 1-2220 nMillhr. Its maximum value was recortled on August and minimum during October and November. The PA and chlorophyll u v~luess howed high correlation coefficent (0.69). and the values of specific astivity was highest during Winter, 45-12\ulcorner nMihripg chl. a, and lowest on October. 2 nMl hripg chl. 11. The values of PA and bioavailable dissolved inorganic phosphate (DIP) showed negative correlation in surface water. During destratification period (Oct. and Nov. 1990). the values of PA were about 11240 times lower than those during August albeit high concentration of chlorophyll a (1.7- 7.2 mglm'). Such results seem to be cause'd by DIP supply from the metalimnion. hypolimnion ant1 sediment. With these results. phosphate was sufficient to sustain the biomass in Autumn. and internal loading of phosphate shoultl accelerate the eutrophication in Lake Soyang.
Kim, Jin;Yook, Jong-In;Park, Kwang-Kyun;Lee, Eun-Ha;Jung, So-Young;Joon, Yin-Liu;Kyung, Chul-Hong;Kim, Ju;Chung, Won-Yoon
Environmental Mutagens and Carcinogens
/
v.19
no.1
/
pp.46-55
/
1999
To develop a chemopreventive strategy based on the different stages of premalignant lesions, we hypothesized that the inhibitory effect of chemopreventive agents is influenced by different promotion stages during carcinogenesis. DMBA-TPA-induced skin carcinogenesis was used with ICR mice and chlorophyllin (CHL) was applied as a chemopreventive agent. In vitro assay was performed with Salmonella typhi. TA100 to observe any anti-mutagenic activity of CHL against DMBA. Pre-initiation and pre-promotion effects of CHL were observed by CHL treatment before initiation and before promotion. To evaluate the inhibitory effect at different promotion stages, each group was divided into three subgroups after TPA promotion for 6, 18 and 24 weeks, respectively ; the first subgroup was immediately sacrificed after termination of TPA, the second subgroup was treated with CHL, and the third subgroup was sacrificed 8 weeks after termination of TPA without CHL treatment. The degrees of epithelial dysplasia, papilloma formation, and invasive carcinoma were observed histologically, and GST-Pi expression was observed immunohistochemically. ODC mRNA level was analyzed by reverse transcriptase-polymerase chain reaction. Results showed : CHL dose-dependently inhibited the mutation of Salmonella typhi. TA100; the incidence of epithelial dysplasia and papilloma formation was lower in pre-initiation and pre-promotion CHL-treated mice than DMBA-TPA-treated mice; no invasive carcinoma developed in pre-initiation CHL-treated groups, while 67% of DMBA-TPA treated mice had carcinomas; GST-Pi expression decreased when CHL was treated before initiation and before promotion; and when CHL was treated after termination of TPA application at 18 and 24- week-TPA promotion stages, respectively, the incidence of epithelial dysplasia and papilloma was markedly reduced compared to non-treated groups. When comparing the incidence of epithelial dysplasia and papilloma between the immediately-sacrificed subgroup and the non-treated group with a waiting period, we speculated that the 18-week-TPA promotion stage might belong to the promoter-independent progression stage. At the 18-week-TPA promotion stage, the level of ODC mRNA in the CHL-treated group was clearly reduced to the level of normal tissue. Taking these results together, CHL showed both anti-initiation and anti-promotion effects, while the inhibitory effect of CHL was prominent in the 18-week-TPA promotion stage. However, CHL seems to be incapable of completely blocking the progression in the 24-week-TPA promotion stage.
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