BACKGROUND Chronic nonhealing wounds,such as diabetic foot ulcer(DFU),suffer from delayed healing.Identifying effective biomarkers or targets is crucial for managing these refractory wounds.While N7-methylguanosine(m7...BACKGROUND Chronic nonhealing wounds,such as diabetic foot ulcer(DFU),suffer from delayed healing.Identifying effective biomarkers or targets is crucial for managing these refractory wounds.While N7-methylguanosine(m7G)methylation is important in RNA modification,its connection to chronic nonhealing wounds is poorly understood.AIM To assess the potential m7G biomarkers in DFU and their underlying molecular mechanisms.METHODS Differential expression analysis and weighted gene coexpression network analysis identified key genes in DFU.Hub genes were determined through m7G-DFU intersection,and gene set enrichment analysis was conducted.Diagnostic potential of hub genes was assessed using receiver operating characteristic curves.The hub gene’s expression(decapping scavenger enzyme,DCPS)was confirmed using quantitative reverse transcription polymerase chain reaction and immunofluorescence.In vitro,normal human epidermal keratinocyte models were knocked down for DCPS,and the function was assessed through flow cytometry,western blotting,immunofluorescence,Transwell assays,and scratch assays.RESULTS Weighted gene coexpression network analysis and differential expression analysis revealed links between DFU datasets and methylation processes,identifying hub gene DCPS as a candidate biomarker.Notably,its diagnostic value was confirmed with a test set and receiver operating characteristic curve,achieving an area under the curve of 0.98 and 0.99.Quantitative reverse transcription polymerase chain reaction and immunofluorescence analyses showed significantly reduced expression of DCPS in the wound skin of DFU patients and streptozotocin-induced diabetic mice,indicating its role as a regulatory factor of m7G in diabetic wounds.Mechanistically,in vitro studies showed that DCPS knockdown significantly reduced cyclin-dependent kinase 6 and cyclin D1 expression,disrupted the epithelial cell cycle,inhibited cell proliferation and migration,and increased apoptosis rates.CONCLUSION DCPS was identified as a promising DFU biomarker and therapeutic target,regulating m7G to affect cell cycle,proliferation,and epithelial cell migration during DFU wound healing.展开更多
The plasticity of stem cells in response to environmental change is critical for multicellular organisms.Here,we show that MYB3R-like directly activates the key plant stem-cell regulator WUSCHEL(WUS)by recruiting the ...The plasticity of stem cells in response to environmental change is critical for multicellular organisms.Here,we show that MYB3R-like directly activates the key plant stem-cell regulator WUSCHEL(WUS)by recruiting the methyltransferase ROOT INITIATION DEFECTIVE 2(RID2),which functions in m7G methylation of the 5′cap of WUS mRNA to protect it from degradation.Transcriptomic and molecular analyses showed that protein-folding genes are repressed by WUS to maintain precise protein synthesis in stem cells by preventing the reuse of misfolded proteins.Interestingly,we found that upon heat stress,the MYB3R-like/RID2 module is repressed to reduce WUS transcript abundance through decapping of nascent WUS mRNA.This releases the inhibition of protein-folding capacity in stem cells and protects them from heat shock by eliminating misfolded protein aggregation.Taken together,our results reveal a strategic trade-off whereby plants reduce the accuracy of protein synthesis in exchange for the survival of stem cells at high temperatures.展开更多
Human NUDT16(hNUDT16)is a decapping enzyme initially identified as the human homolog to the Xenopus laevis X29.As a metalloenzyme,hNUDT16 relies on divalent cations for its cap-hydrolysis activity to remove m7 GDP and...Human NUDT16(hNUDT16)is a decapping enzyme initially identified as the human homolog to the Xenopus laevis X29.As a metalloenzyme,hNUDT16 relies on divalent cations for its cap-hydrolysis activity to remove m7 GDP and m227GDP from RNAs.Metal also determines substrate specificity of the enzyme.So far,only U8 small nucleolar RNA(snoRNA)has been identified as the substrate of hNUDT16 in the presence of Mg2+.Here we demonstrate that besides U8,hNUDT16 can also actively cleave the m7 GDP cap from mRNAs in the presence of Mg2+or Mn2+.We further show that hNUDT16 does not preferentially recognize U8 or mRNA substrates by our cross-inhibition and quantitative decapping assays.In addition,our mutagenesis analysis identifies several key residues involved in hydrolysis and confirms the key role of the REXXEE motif in catalysis.Finally an investigation into the subcellular localization of hNUDT16 revealed its abundance in both cytoplasm and nucleus.These findings extend the substrate spectrum of hNUDT16 beyond snoRNAs to also include mRNA,demonstrating the pleiotropic decapping activity of hNUDT16.展开更多
The objective of the study was to establish the effect of formic acid on varroa(Varroa destructor),inside the capped brood cells,artificially decapped.The experiments were carried out in 2017-2018 on honeybee colonies...The objective of the study was to establish the effect of formic acid on varroa(Varroa destructor),inside the capped brood cells,artificially decapped.The experiments were carried out in 2017-2018 on honeybee colonies infested with varroa(V.destructor),in a research apiary belonging to the Institute for Beekeeping Research and Development in Bucharest.The decapping method in the present researches used the decapping fork to scrape the capped comb,without affecting the brood,in order to open it for an effective treatment.The combined treatment method was applied on honeybee colonies as a whole,as well as on brood combs,without bees,put in a special treatment box.The researches were focused on establishing the mortality level of various stages of varroa in artificially decapped brood,in normal colony and separately,as well as to make observations on the effect of formic acid on viability of capped bee brood,artificially decapped.The results show a high mortality of varroa,especially the protonymphs and deutonymphs stages(over 80%).The main conclusion is that the brood decapping method combined with formic acid treatment could be a useful technique to control varroa infestation,both in brood and honeybees,shortening strongly the treatment duration as compared to the usual treatments,increasing the efficacy of treatment by cutting the life cycle of varroa in brood.展开更多
基金Supported by National Natural Science Foundation of China,No.82370903Noncommunicable Chronic Diseases-National Science and Technology Major Project,No.2023ZD0509400 and No.2023ZD0509402+1 种基金2023 Key Disciplines on Public Health Construction of Chongqing,the Natural Science Foundation of Chongqing Municipal Science and Technology Bureau,No.cstc2024ycjh-bgzxm0014Major Project of Science and Technology Research Program of Chongqing Education Commission of China,No.KJZD-M202400102.
文摘BACKGROUND Chronic nonhealing wounds,such as diabetic foot ulcer(DFU),suffer from delayed healing.Identifying effective biomarkers or targets is crucial for managing these refractory wounds.While N7-methylguanosine(m7G)methylation is important in RNA modification,its connection to chronic nonhealing wounds is poorly understood.AIM To assess the potential m7G biomarkers in DFU and their underlying molecular mechanisms.METHODS Differential expression analysis and weighted gene coexpression network analysis identified key genes in DFU.Hub genes were determined through m7G-DFU intersection,and gene set enrichment analysis was conducted.Diagnostic potential of hub genes was assessed using receiver operating characteristic curves.The hub gene’s expression(decapping scavenger enzyme,DCPS)was confirmed using quantitative reverse transcription polymerase chain reaction and immunofluorescence.In vitro,normal human epidermal keratinocyte models were knocked down for DCPS,and the function was assessed through flow cytometry,western blotting,immunofluorescence,Transwell assays,and scratch assays.RESULTS Weighted gene coexpression network analysis and differential expression analysis revealed links between DFU datasets and methylation processes,identifying hub gene DCPS as a candidate biomarker.Notably,its diagnostic value was confirmed with a test set and receiver operating characteristic curve,achieving an area under the curve of 0.98 and 0.99.Quantitative reverse transcription polymerase chain reaction and immunofluorescence analyses showed significantly reduced expression of DCPS in the wound skin of DFU patients and streptozotocin-induced diabetic mice,indicating its role as a regulatory factor of m7G in diabetic wounds.Mechanistically,in vitro studies showed that DCPS knockdown significantly reduced cyclin-dependent kinase 6 and cyclin D1 expression,disrupted the epithelial cell cycle,inhibited cell proliferation and migration,and increased apoptosis rates.CONCLUSION DCPS was identified as a promising DFU biomarker and therapeutic target,regulating m7G to affect cell cycle,proliferation,and epithelial cell migration during DFU wound healing.
基金National Natural Science Foundation of China(grant nos.32321001 and 32130009 to Z.Z.)University of Science and Technology of China Research Funds of the Double First-Class Initiative(grant no.YD9100002025 to Z.Z.).
文摘The plasticity of stem cells in response to environmental change is critical for multicellular organisms.Here,we show that MYB3R-like directly activates the key plant stem-cell regulator WUSCHEL(WUS)by recruiting the methyltransferase ROOT INITIATION DEFECTIVE 2(RID2),which functions in m7G methylation of the 5′cap of WUS mRNA to protect it from degradation.Transcriptomic and molecular analyses showed that protein-folding genes are repressed by WUS to maintain precise protein synthesis in stem cells by preventing the reuse of misfolded proteins.Interestingly,we found that upon heat stress,the MYB3R-like/RID2 module is repressed to reduce WUS transcript abundance through decapping of nascent WUS mRNA.This releases the inhibition of protein-folding capacity in stem cells and protects them from heat shock by eliminating misfolded protein aggregation.Taken together,our results reveal a strategic trade-off whereby plants reduce the accuracy of protein synthesis in exchange for the survival of stem cells at high temperatures.
基金the Natural Science Foundation of China(No.30870118)。
文摘Human NUDT16(hNUDT16)is a decapping enzyme initially identified as the human homolog to the Xenopus laevis X29.As a metalloenzyme,hNUDT16 relies on divalent cations for its cap-hydrolysis activity to remove m7 GDP and m227GDP from RNAs.Metal also determines substrate specificity of the enzyme.So far,only U8 small nucleolar RNA(snoRNA)has been identified as the substrate of hNUDT16 in the presence of Mg2+.Here we demonstrate that besides U8,hNUDT16 can also actively cleave the m7 GDP cap from mRNAs in the presence of Mg2+or Mn2+.We further show that hNUDT16 does not preferentially recognize U8 or mRNA substrates by our cross-inhibition and quantitative decapping assays.In addition,our mutagenesis analysis identifies several key residues involved in hydrolysis and confirms the key role of the REXXEE motif in catalysis.Finally an investigation into the subcellular localization of hNUDT16 revealed its abundance in both cytoplasm and nucleus.These findings extend the substrate spectrum of hNUDT16 beyond snoRNAs to also include mRNA,demonstrating the pleiotropic decapping activity of hNUDT16.
文摘The objective of the study was to establish the effect of formic acid on varroa(Varroa destructor),inside the capped brood cells,artificially decapped.The experiments were carried out in 2017-2018 on honeybee colonies infested with varroa(V.destructor),in a research apiary belonging to the Institute for Beekeeping Research and Development in Bucharest.The decapping method in the present researches used the decapping fork to scrape the capped comb,without affecting the brood,in order to open it for an effective treatment.The combined treatment method was applied on honeybee colonies as a whole,as well as on brood combs,without bees,put in a special treatment box.The researches were focused on establishing the mortality level of various stages of varroa in artificially decapped brood,in normal colony and separately,as well as to make observations on the effect of formic acid on viability of capped bee brood,artificially decapped.The results show a high mortality of varroa,especially the protonymphs and deutonymphs stages(over 80%).The main conclusion is that the brood decapping method combined with formic acid treatment could be a useful technique to control varroa infestation,both in brood and honeybees,shortening strongly the treatment duration as compared to the usual treatments,increasing the efficacy of treatment by cutting the life cycle of varroa in brood.
