AIM:To examine the methylation status of the promoter region of the checkpoint with forkhead-associated and ring fi nger(CHFR) and microsatellite mutator status in 59 primary gastric cancers.METHODS:We investigated th...AIM:To examine the methylation status of the promoter region of the checkpoint with forkhead-associated and ring fi nger(CHFR) and microsatellite mutator status in 59 primary gastric cancers.METHODS:We investigated the promoter methylation of CHFR in 59 cases of gastric cancer using methylation-specifi c PCR.Five microsatellite loci were analyzed using high-intensity microsatellite analysis reported previously, and p53 gene mutations were investigated by direct sequencing.RESULTS:Twenty cases(33.9%) showed promoter methylation and no relation was observed with the clinicopathological factors.We found that the promoter methylation of CHFR was frequently accompanied with microsatellite instability(MIN).Seven of 20(35.0%) cases showed MIN in hypermethylation of the CHFR tumor, while three of 39(7.7%) cases showed MIN in the non-methylated CHFR tumor(P < 0.01).However, we failed to fi nd any relationship between CHFR methylation and p53 mutation status.CONCLUSION:The coordinated loss of both the mitotic check point function and mismatch repair system suggests the potential to overcome the cell cycle check point, which may lead to an accumulation of mutations.However, the p53 mutation was not related to hypermethylation of the CHFR promoter and MIN, which indicates that an abnormality in p53 occurs as an independent process from the mismatch repair deficiency in carcinogenesis.展开更多
Dawdle （DDL） is a microRNA processing protein essential for the development of Arabidopsis. DDL contains a putative nuclear localization signal at its amino-terminus and forkhead-associated （FHA） domain at the car...Dawdle （DDL） is a microRNA processing protein essential for the development of Arabidopsis. DDL contains a putative nuclear localization signal at its amino-terminus and forkhead-associated （FHA） domain at the carboxyl-termi- nus. Here, we report the crystal structure of the FHA domain of Arabidopsis Dawdle, determined by multiple-wavelength anomalous dispersion method at 1.7-A resolution. DDL FHA structure displays a seven-stranded 13-sandwich architec- ture that contains a unique structural motif comprising two long anti-parallel strands. Strikingly, crystal packing of the DDL FHA domain reveals that a glutamate residue from the symmetry-related DDL FHA domain, a structural mimic of the phospho-threonine, is specifically recognized by the structurally conserved phospho-threonine binding cleft. Consistently with the structural observations, co-immuno-precipitation experiments performed in Nicotiana benthami- ana show that the DDL FHA domain co-immuno-precipitates with DCL1 fragments containing the predicted pThr＋3（lle/ Val/Leu/Asp） motif. Taken together, we count the recognition of the target residue by the canonical binding cleft of the DDL FHA domain as the key molecular event to instate FHA domain-mediated protein-protein interaction in plant miRNA processing.展开更多
Peroxisomes are ubiquitous and dynamic eukaryotic organelles capable of altering their abun- dance in response to environmental and developmental cues, yet the regulatory mechanism of plant peroxisome division/prolife...Peroxisomes are ubiquitous and dynamic eukaryotic organelles capable of altering their abun- dance in response to environmental and developmental cues, yet the regulatory mechanism of plant peroxisome division/proliferation is unclear. To identify transcriptional regulators of the peroxisome division factor gene PEX 11b, we performed a nuclear pull-clown experiment and identified Arabidopsis Forkhead- Associated Domain Protein 3 （FHA3） as a novel protein that binds to the promoter of PEX 11b. Our data supported the conclusion that, in contrast to the previously identified HY5 HOMOLOG （HYH） protein that promotes the transcription of PEX 11b, FHA3 is a negative regulator of PEX 11b expression and peroxisome division.展开更多
文摘AIM:To examine the methylation status of the promoter region of the checkpoint with forkhead-associated and ring fi nger(CHFR) and microsatellite mutator status in 59 primary gastric cancers.METHODS:We investigated the promoter methylation of CHFR in 59 cases of gastric cancer using methylation-specifi c PCR.Five microsatellite loci were analyzed using high-intensity microsatellite analysis reported previously, and p53 gene mutations were investigated by direct sequencing.RESULTS:Twenty cases(33.9%) showed promoter methylation and no relation was observed with the clinicopathological factors.We found that the promoter methylation of CHFR was frequently accompanied with microsatellite instability(MIN).Seven of 20(35.0%) cases showed MIN in hypermethylation of the CHFR tumor, while three of 39(7.7%) cases showed MIN in the non-methylated CHFR tumor(P < 0.01).However, we failed to fi nd any relationship between CHFR methylation and p53 mutation status.CONCLUSION:The coordinated loss of both the mitotic check point function and mismatch repair system suggests the potential to overcome the cell cycle check point, which may lead to an accumulation of mutations.However, the p53 mutation was not related to hypermethylation of the CHFR promoter and MIN, which indicates that an abnormality in p53 occurs as an independent process from the mismatch repair deficiency in carcinogenesis.
文摘Dawdle （DDL） is a microRNA processing protein essential for the development of Arabidopsis. DDL contains a putative nuclear localization signal at its amino-terminus and forkhead-associated （FHA） domain at the carboxyl-termi- nus. Here, we report the crystal structure of the FHA domain of Arabidopsis Dawdle, determined by multiple-wavelength anomalous dispersion method at 1.7-A resolution. DDL FHA structure displays a seven-stranded 13-sandwich architec- ture that contains a unique structural motif comprising two long anti-parallel strands. Strikingly, crystal packing of the DDL FHA domain reveals that a glutamate residue from the symmetry-related DDL FHA domain, a structural mimic of the phospho-threonine, is specifically recognized by the structurally conserved phospho-threonine binding cleft. Consistently with the structural observations, co-immuno-precipitation experiments performed in Nicotiana benthami- ana show that the DDL FHA domain co-immuno-precipitates with DCL1 fragments containing the predicted pThr＋3（lle/ Val/Leu/Asp） motif. Taken together, we count the recognition of the target residue by the canonical binding cleft of the DDL FHA domain as the key molecular event to instate FHA domain-mediated protein-protein interaction in plant miRNA processing.
基金the Arabidopsis Biological Resource Center(ABRC)for providing seeds of T-DNA insertion mutant fha3-1(Salk_095831)supported by grants from the National Science Foundation(MCB 0618335,MCB 1330441)the Chemical Sciences,Geosciences and Biosciences Division,Office of Basic Energy Sciences,Office of Science,U.S. Department of Energy(DE-FG02-91ER20021)to J.H
文摘Peroxisomes are ubiquitous and dynamic eukaryotic organelles capable of altering their abun- dance in response to environmental and developmental cues, yet the regulatory mechanism of plant peroxisome division/proliferation is unclear. To identify transcriptional regulators of the peroxisome division factor gene PEX 11b, we performed a nuclear pull-clown experiment and identified Arabidopsis Forkhead- Associated Domain Protein 3 （FHA3） as a novel protein that binds to the promoter of PEX 11b. Our data supported the conclusion that, in contrast to the previously identified HY5 HOMOLOG （HYH） protein that promotes the transcription of PEX 11b, FHA3 is a negative regulator of PEX 11b expression and peroxisome division.
