Glutamate dehydrogenase regulates crop development, growth, and biomass yield through its synthesis of non-genetic code-based RNA. Understanding the mechanism of GDH-synthesized RNA enzyme would enhance the agricultur...Glutamate dehydrogenase regulates crop development, growth, and biomass yield through its synthesis of non-genetic code-based RNA. Understanding the mechanism of GDH-synthesized RNA enzyme would enhance the agriculture innovation capacity of the more than a billion urban gardeners, smallholder, and limited resources indigenous farmers. Different metabolic variants were prepared by treating peanut growing on healthy soil with stoichiometric mixes of mineral salt solutions. Peanut GDH charge isomers were purified to homogeneity by electrophoresis, and made to synthesize RNA enzyme. Peanut total RNA was 5’-end labeled with [γ-32P]ATP and made to react as substrate in vitro with GDH-synthesized RNA from another metabolic variant of peanut. Agarose, and polyacrylamide gel electrophoresis of the reaction products showed that tRNA, rRNA, and most of the mRNAs were degraded to mononucleotides, but total RNAs that were not mixed with GDH-synthesized RNAs were not degraded. When the non-homologous sequence sections of the GDH-synthesized RNA were clipped out, the homologous sections failed to produce Northern bands with peanut total RNA. Therefore, the non-homologous sequence sections served to identify, position, and align the GDH-synthesized RNA to its target total RNA site independent of genetic code;the degradation of total RNA being via non-canonical base alignments in the enzyme-substrate complex, followed by electromagnetic destruction of the total RNA, the less stable of the two kinds of RNA. This is the science-based corner stone that buttresses the crop production efforts of limited resources farmers because GDH-synthesized RNAs quickly degrade superfluous total RNA of the crop in response to the soil mineral nutrient deficiencies thereby minimizing wastage of metabolic energy in the synthesis of unnecessary protein enzymes while optimizing biomass metabolism, crop growth, and maximum crop yields. In vitro hydrolysis of total RNA by GDH-synthesized RNA is the game changing, prototype, R&D methods for cleansing sick total RNA from cells, tissues, and whole organisms.展开更多
Circular intronic RNAs(ci RNAs) escaping from DBR1 debranching of intron lariats are co-transcriptionally produced from prem RNA splicing, but their turnover and mechanism of action have remained elusive. We report th...Circular intronic RNAs(ci RNAs) escaping from DBR1 debranching of intron lariats are co-transcriptionally produced from prem RNA splicing, but their turnover and mechanism of action have remained elusive. We report that RNase H1 degrades a subgroup of ci RNAs in human cells. Many ci RNAs contain high GC% and tend to form DNA:RNA hybrids(R-loops) for RNase H1 cleavage, a process that appears to promote Pol II transcriptional elongation at ci RNA-producing loci. One ci RNA, ciankrd52, shows a stronger ability of R-loop formation than that of its cognate pre-m RNA by maintaining a locally open RNA structure in vitro. This allows the release of pre-m RNA from R-loops by ci-ankrd52 replacement and subsequent ci RNA removal via RNase H1 for efficient transcriptional elongation. We propose that such an R-loop dependent ci RNA degradation likely represents a mechanism that on one hand limits ci RNA accumulation by recruiting RNase H1 and on the other hand resolves Rloops for transcriptional elongation at some GC-rich ci RNA-producing loci.展开更多
文摘Glutamate dehydrogenase regulates crop development, growth, and biomass yield through its synthesis of non-genetic code-based RNA. Understanding the mechanism of GDH-synthesized RNA enzyme would enhance the agriculture innovation capacity of the more than a billion urban gardeners, smallholder, and limited resources indigenous farmers. Different metabolic variants were prepared by treating peanut growing on healthy soil with stoichiometric mixes of mineral salt solutions. Peanut GDH charge isomers were purified to homogeneity by electrophoresis, and made to synthesize RNA enzyme. Peanut total RNA was 5’-end labeled with [γ-32P]ATP and made to react as substrate in vitro with GDH-synthesized RNA from another metabolic variant of peanut. Agarose, and polyacrylamide gel electrophoresis of the reaction products showed that tRNA, rRNA, and most of the mRNAs were degraded to mononucleotides, but total RNAs that were not mixed with GDH-synthesized RNAs were not degraded. When the non-homologous sequence sections of the GDH-synthesized RNA were clipped out, the homologous sections failed to produce Northern bands with peanut total RNA. Therefore, the non-homologous sequence sections served to identify, position, and align the GDH-synthesized RNA to its target total RNA site independent of genetic code;the degradation of total RNA being via non-canonical base alignments in the enzyme-substrate complex, followed by electromagnetic destruction of the total RNA, the less stable of the two kinds of RNA. This is the science-based corner stone that buttresses the crop production efforts of limited resources farmers because GDH-synthesized RNAs quickly degrade superfluous total RNA of the crop in response to the soil mineral nutrient deficiencies thereby minimizing wastage of metabolic energy in the synthesis of unnecessary protein enzymes while optimizing biomass metabolism, crop growth, and maximum crop yields. In vitro hydrolysis of total RNA by GDH-synthesized RNA is the game changing, prototype, R&D methods for cleansing sick total RNA from cells, tissues, and whole organisms.
基金This work was supported by the National Natural Science Foundation of China(NSFC)(91940303,31725009)the HHMI International Program(55008728)to L.-L.C.+2 种基金NSFC(31730111,31925011)to L.Y.Young Elite Scientists Sponsorship Program(2020QNRC001)to X.L.L.-L.C.the support from the XPLORER PRIZE.
文摘Circular intronic RNAs(ci RNAs) escaping from DBR1 debranching of intron lariats are co-transcriptionally produced from prem RNA splicing, but their turnover and mechanism of action have remained elusive. We report that RNase H1 degrades a subgroup of ci RNAs in human cells. Many ci RNAs contain high GC% and tend to form DNA:RNA hybrids(R-loops) for RNase H1 cleavage, a process that appears to promote Pol II transcriptional elongation at ci RNA-producing loci. One ci RNA, ciankrd52, shows a stronger ability of R-loop formation than that of its cognate pre-m RNA by maintaining a locally open RNA structure in vitro. This allows the release of pre-m RNA from R-loops by ci-ankrd52 replacement and subsequent ci RNA removal via RNase H1 for efficient transcriptional elongation. We propose that such an R-loop dependent ci RNA degradation likely represents a mechanism that on one hand limits ci RNA accumulation by recruiting RNase H1 and on the other hand resolves Rloops for transcriptional elongation at some GC-rich ci RNA-producing loci.
