Heat stress can stimulate an increase in body temperature, which is correlated with increased expression of heat shock protein 70 （HSP70） and tumor necrosis factor a （TNFa）. The exact mechanism underlying the HSP7...Heat stress can stimulate an increase in body temperature, which is correlated with increased expression of heat shock protein 70 （HSP70） and tumor necrosis factor a （TNFa）. The exact mechanism underlying the HSP70 and TNFa induction is unclear. Berberine （BBR） can significantly inhibit the temperature rise caused by heat stress, but the mechanism responsible for the BBR effect on HSP70 and TNFa signaling has not been investigated. The aim of the present study was to explore the relationship between the expression of HSP70 and TNFa and the effects of BBR under heat conditions, using in vivo and in vitro models. The expression levels of HSP70 and YNFa were determined using RT-PCR and Western blotting analyses. The results showed that the levels of HSP70 and TNFa were ap-regulated under heat conditions （40 ~C）. HSP70 acted as a chaperone to maintain TNFa homeostasis with rising the temperature, but knockdown of HSP70 could not down-regulate the level of TNFa. Furthermore, TNFa could not influence the expression of HSP70 under aormal and heat conditions. BBR targeted both HSP70 and TNFa by suppressing their gene transcription, thereby decreasing body temperature under heat conditions. In conclusion, BBR has a potential to be developed as a therapeutic strategy for suppressing the thermal effects in hot environments.展开更多
Myostatin(MSTN) is one of the key factors regulating myogenesis. Because of its role as a negative regulator of muscle mass deposition, much interest has been given to its protein and, in recent years, several studies...Myostatin(MSTN) is one of the key factors regulating myogenesis. Because of its role as a negative regulator of muscle mass deposition, much interest has been given to its protein and, in recent years, several studies have analysed MSTN gene regulation. This review discusses the MSTN gene promoter, focusing on its structure in several animal species, both vertebrate and invertebrate. We report the important binding sites considering their degree of phylogenetic conservation and roles they play in the promoter activity. Finally, we discuss recent studies focusing on MSTN gene regulation via promoter manipulation and the potential applications they have both in medicine and agriculture.展开更多
Short-chain fatty acids are important nutrients that regulate milk fat synthesis.They regulate milk syn-thesis via the sterol regulatory element binding protein 1(SREBP1)pathway;however,the details are still unknown.H...Short-chain fatty acids are important nutrients that regulate milk fat synthesis.They regulate milk syn-thesis via the sterol regulatory element binding protein 1(SREBP1)pathway;however,the details are still unknown.Here,the regulation and mechanism of sodium acetate(SA)in milk fat synthesis in bovine mammary epithelial cells(BMECs)were assessed.BMECs were treated with SA supplementation(SAþ)or without SA supplementation(SA-),and milk fat synthesis and activation of the SREBP1 pathway were increased(P=0.0045;P=0.0042)by SAþand decreased(P=0.0068;P=0.0031)by SA-,respectively.Overexpression or inhibition of SREBP1 demonstrated that SA promoted milk fat synthesis(P=0.0045)via the SREBP1 pathway.Overexpression or inhibition of TATA element modulatory factor 1(TMF1)demon-strated that TMF1 suppressed activation of the SREBP1 pathway(P=0.0001)and milk fat synthesis(P=0.0022)activated by SAþ.Overexpression or inhibition of TMF1 and SREBP1 showed that TMF1 suppressed milk fat synthesis(P=0.0073)through the SREBP1 pathway.Coimmunoprecipitation analysis revealed that TMF1 interacted with SREBP1 in the cytoplasm and suppressed the nuclear localization of SREBP1(P=0.0066).The absence or presence of SA demonstrated that SA inhibited the expression of TMF1(P=0.0002)and the interaction between TMF1 and SREBP1(P=0.0001).Collectively,our research sug-gested that TMF1 was a new negative regulator of milk fat synthesis.In BMECs,SA promoted the SREBP1 pathway and milk fat synthesis by suppressing TMF1.This study enhances the current understanding of the regulation of milk fat synthesis and provides new scientific data for the regulation of milk fat synthesis.展开更多
The importance of microRNA （miRNA） at the post-transcriptional regulation level has recently been recognized in both animals and plants. In recent years, many studies focused on miRNA target identification and funct...The importance of microRNA （miRNA） at the post-transcriptional regulation level has recently been recognized in both animals and plants. In recent years, many studies focused on miRNA target identification and functional analysis. However, little is known about the transcription and regulation of miRNAs themselves. In this study, the transcription start sites （TSSs） for 11 miRNA primary transcripts of soybean from 11 miRNA loci （of 50 loci tested） were cloned by a 5＂ rapid amplification of cDNA ends （5＂ RACE） procedure using total RNA from 30-d-old seedlings. The features consistent with a RNA polymerase II mechanism of transcription were found among these miRNA loci. A position weight matrix algorithm was used to identify conserved motifs in miRNA core promoter regions. A canonical TATA box motif was identified upstream of the major start site at 8 （76%） of the mapped miRNA loci. Several cis-acting elements were predicted in the 2 kb 5＂ to the TSSs. Potential spatial and temporal expression patterns of the miRNAs were found. The target genes for these miRNAs were also predicted and further elucidated for the potential function of the miRNAs. This research provides a molecular basis to explore regulatory mechanisms of miRNA expression, and a way to understand miRNA-mediated regulatory pathways and networks in soybean.展开更多
In polyglutamine(PolyQ)diseases,mutant proteins cause not only neurological problems but also peripheral tissue abnormalities.Among all systemic damages,skeletal muscle dystrophy is the severest.Previously by studying...In polyglutamine(PolyQ)diseases,mutant proteins cause not only neurological problems but also peripheral tissue abnormalities.Among all systemic damages,skeletal muscle dystrophy is the severest.Previously by studying knock-in(KI)mouse models of spinal cerebellar ataxia 17(SCA17),it was found that mutant TATA box binding protein(TBP)decreases its interaction with myogenic differentiation antigen,thus reducing the expression of skeletal muscle structural proteins and resulting in muscle degeneration.In this paper,the role of mutant TBP in myogenesis was investigated.Single myofibers were isolated from tibialis anterior muscles of wild type(WT)and SCA17KI mice.The 1TBP18 staining confirmed the expression of mutant TBP in muscle satellite cells in SCA17Ki mice.In the BaCl2-induced TA muscle injury,H&E cross-section staining showed no significant change in myofibril size before and after BaCl2 treatment,and there was no significant difference in centralized nuclei between WT and SCA17KI mice,suggesting that mutant TBP had no significant effect on muscle regeneration.In the cultured primary myoblasts from WT and SCA17KI mice in vitro,representative BrdU immunostaining showed no significant difference in proliferation of muscle satellite cells.The primary myoblasts were then induced to differentiate and immunostained for eMyHC,and the staining showed there was no significant difference in differentiation of primary myoblasts between WT and SCA1KI mice.Our findings confirmed that mutant TBP had no significant effect on myogenesis.展开更多
The nonlinear Poisson-Boltzmann predictions of the salt-dependent association of proteins to DNA,SKpred,are fairly insensitive to the choice of atomic charges,radii,interior dielectric constant and treatment of the bo...The nonlinear Poisson-Boltzmann predictions of the salt-dependent association of proteins to DNA,SKpred,are fairly insensitive to the choice of atomic charges,radii,interior dielectric constant and treatment of the boundary between a biomolecule and the solvent.In this study we show that the SKpred is highly correlated with the conformational adaptability of the partners involved in the biomolecular binding process.This is demonstrated for the wild-type and mutant forms of the archaeon Pyrococcus woesi TATA-binding protein(PwTBP)in complex with DNA,on which we performed molecular mechanics energy minimizations with different protocols,and molecular dynamics simulations and then computed the SKpred on the resulting structures.It was found that the inter-molecular non bonded force field energy between the DNA and protein correlates linearly and significantly well with the SKpred.This correlation encompasses the wild-type and mutant variants of the PwTBP and provides us with a quick way to estimate the SKpred from a large ensemble of structures generated with Molecular Dynamics or Monte Carlo simulations.The corresponding experimental SKobs should also correlate with the inter-molecular non bonded force field energy between the protein and DNA,given that the underlying mechanisms in binding and salt-dependent effects are in fact the main contributors in the association of proteins/peptides to nucleic acids.We show that it is possible to fit experiments versus the inter-molecular non bonded force field energy between the protein and DNA,and use this relation to predict the SKobs in absolute numbers.Thus,we present two novel approaches to estimate both the SKpred and the SKobs for in silico modelled PwTBP novel mutants and even for TBPs from other organisms.This is a simple but powerful tool to suggest new experiments on the TBP-DNA type of macromolecular assemblies.We conclude by suggesting some mutants and a possible biological interpretation of how changes in solvent salinity affect the binding of proteins to DNA.展开更多
基金supported by the National Natural Science Foundation of China(Nos.81374006,90713043 and 81073092)
文摘Heat stress can stimulate an increase in body temperature, which is correlated with increased expression of heat shock protein 70 （HSP70） and tumor necrosis factor a （TNFa）. The exact mechanism underlying the HSP70 and TNFa induction is unclear. Berberine （BBR） can significantly inhibit the temperature rise caused by heat stress, but the mechanism responsible for the BBR effect on HSP70 and TNFa signaling has not been investigated. The aim of the present study was to explore the relationship between the expression of HSP70 and TNFa and the effects of BBR under heat conditions, using in vivo and in vitro models. The expression levels of HSP70 and YNFa were determined using RT-PCR and Western blotting analyses. The results showed that the levels of HSP70 and TNFa were ap-regulated under heat conditions （40 ~C）. HSP70 acted as a chaperone to maintain TNFa homeostasis with rising the temperature, but knockdown of HSP70 could not down-regulate the level of TNFa. Furthermore, TNFa could not influence the expression of HSP70 under aormal and heat conditions. BBR targeted both HSP70 and TNFa by suppressing their gene transcription, thereby decreasing body temperature under heat conditions. In conclusion, BBR has a potential to be developed as a therapeutic strategy for suppressing the thermal effects in hot environments.
文摘Myostatin(MSTN) is one of the key factors regulating myogenesis. Because of its role as a negative regulator of muscle mass deposition, much interest has been given to its protein and, in recent years, several studies have analysed MSTN gene regulation. This review discusses the MSTN gene promoter, focusing on its structure in several animal species, both vertebrate and invertebrate. We report the important binding sites considering their degree of phylogenetic conservation and roles they play in the promoter activity. Finally, we discuss recent studies focusing on MSTN gene regulation via promoter manipulation and the potential applications they have both in medicine and agriculture.
基金supported by China Postdoctoral Science Foundation funded project(2019M662971)The Basic Scientific Research Operating Expenses of Higher Education Institutions of Heilongjiang Province(No.2020-KYYWF-0283).
文摘Short-chain fatty acids are important nutrients that regulate milk fat synthesis.They regulate milk syn-thesis via the sterol regulatory element binding protein 1(SREBP1)pathway;however,the details are still unknown.Here,the regulation and mechanism of sodium acetate(SA)in milk fat synthesis in bovine mammary epithelial cells(BMECs)were assessed.BMECs were treated with SA supplementation(SAþ)or without SA supplementation(SA-),and milk fat synthesis and activation of the SREBP1 pathway were increased(P=0.0045;P=0.0042)by SAþand decreased(P=0.0068;P=0.0031)by SA-,respectively.Overexpression or inhibition of SREBP1 demonstrated that SA promoted milk fat synthesis(P=0.0045)via the SREBP1 pathway.Overexpression or inhibition of TATA element modulatory factor 1(TMF1)demon-strated that TMF1 suppressed activation of the SREBP1 pathway(P=0.0001)and milk fat synthesis(P=0.0022)activated by SAþ.Overexpression or inhibition of TMF1 and SREBP1 showed that TMF1 suppressed milk fat synthesis(P=0.0073)through the SREBP1 pathway.Coimmunoprecipitation analysis revealed that TMF1 interacted with SREBP1 in the cytoplasm and suppressed the nuclear localization of SREBP1(P=0.0066).The absence or presence of SA demonstrated that SA inhibited the expression of TMF1(P=0.0002)and the interaction between TMF1 and SREBP1(P=0.0001).Collectively,our research sug-gested that TMF1 was a new negative regulator of milk fat synthesis.In BMECs,SA promoted the SREBP1 pathway and milk fat synthesis by suppressing TMF1.This study enhances the current understanding of the regulation of milk fat synthesis and provides new scientific data for the regulation of milk fat synthesis.
基金supported by the National High-Tech R&D Program of China (2006AA10Z1F1)the National Core Soybean Genetic Engineering Project, China(2011ZX08004-002)+3 种基金the National Natural Science Foundation of China (60932008, 30971810)the National Basic Research Program of China (2009CB118400)the Ministry of Education Innovation Team of Soybean Molecular Design,Chinathe Innovation Team of the Education Bureau of Heilongjiang Province, China
文摘The importance of microRNA （miRNA） at the post-transcriptional regulation level has recently been recognized in both animals and plants. In recent years, many studies focused on miRNA target identification and functional analysis. However, little is known about the transcription and regulation of miRNAs themselves. In this study, the transcription start sites （TSSs） for 11 miRNA primary transcripts of soybean from 11 miRNA loci （of 50 loci tested） were cloned by a 5＂ rapid amplification of cDNA ends （5＂ RACE） procedure using total RNA from 30-d-old seedlings. The features consistent with a RNA polymerase II mechanism of transcription were found among these miRNA loci. A position weight matrix algorithm was used to identify conserved motifs in miRNA core promoter regions. A canonical TATA box motif was identified upstream of the major start site at 8 （76%） of the mapped miRNA loci. Several cis-acting elements were predicted in the 2 kb 5＂ to the TSSs. Potential spatial and temporal expression patterns of the miRNAs were found. The target genes for these miRNAs were also predicted and further elucidated for the potential function of the miRNAs. This research provides a molecular basis to explore regulatory mechanisms of miRNA expression, and a way to understand miRNA-mediated regulatory pathways and networks in soybean.
基金This project was supported by grants from the fundamental Research Funds for the Central Universities(No.2019kfyXKJC075)National Key R&D Program of China(No.2017YFC1310000)National Natural Science Foundation of China(No.81671064,and No.81371222).
文摘In polyglutamine(PolyQ)diseases,mutant proteins cause not only neurological problems but also peripheral tissue abnormalities.Among all systemic damages,skeletal muscle dystrophy is the severest.Previously by studying knock-in(KI)mouse models of spinal cerebellar ataxia 17(SCA17),it was found that mutant TATA box binding protein(TBP)decreases its interaction with myogenic differentiation antigen,thus reducing the expression of skeletal muscle structural proteins and resulting in muscle degeneration.In this paper,the role of mutant TBP in myogenesis was investigated.Single myofibers were isolated from tibialis anterior muscles of wild type(WT)and SCA17KI mice.The 1TBP18 staining confirmed the expression of mutant TBP in muscle satellite cells in SCA17Ki mice.In the BaCl2-induced TA muscle injury,H&E cross-section staining showed no significant change in myofibril size before and after BaCl2 treatment,and there was no significant difference in centralized nuclei between WT and SCA17KI mice,suggesting that mutant TBP had no significant effect on muscle regeneration.In the cultured primary myoblasts from WT and SCA17KI mice in vitro,representative BrdU immunostaining showed no significant difference in proliferation of muscle satellite cells.The primary myoblasts were then induced to differentiate and immunostained for eMyHC,and the staining showed there was no significant difference in differentiation of primary myoblasts between WT and SCA1KI mice.Our findings confirmed that mutant TBP had no significant effect on myogenesis.
文摘The nonlinear Poisson-Boltzmann predictions of the salt-dependent association of proteins to DNA,SKpred,are fairly insensitive to the choice of atomic charges,radii,interior dielectric constant and treatment of the boundary between a biomolecule and the solvent.In this study we show that the SKpred is highly correlated with the conformational adaptability of the partners involved in the biomolecular binding process.This is demonstrated for the wild-type and mutant forms of the archaeon Pyrococcus woesi TATA-binding protein(PwTBP)in complex with DNA,on which we performed molecular mechanics energy minimizations with different protocols,and molecular dynamics simulations and then computed the SKpred on the resulting structures.It was found that the inter-molecular non bonded force field energy between the DNA and protein correlates linearly and significantly well with the SKpred.This correlation encompasses the wild-type and mutant variants of the PwTBP and provides us with a quick way to estimate the SKpred from a large ensemble of structures generated with Molecular Dynamics or Monte Carlo simulations.The corresponding experimental SKobs should also correlate with the inter-molecular non bonded force field energy between the protein and DNA,given that the underlying mechanisms in binding and salt-dependent effects are in fact the main contributors in the association of proteins/peptides to nucleic acids.We show that it is possible to fit experiments versus the inter-molecular non bonded force field energy between the protein and DNA,and use this relation to predict the SKobs in absolute numbers.Thus,we present two novel approaches to estimate both the SKpred and the SKobs for in silico modelled PwTBP novel mutants and even for TBPs from other organisms.This is a simple but powerful tool to suggest new experiments on the TBP-DNA type of macromolecular assemblies.We conclude by suggesting some mutants and a possible biological interpretation of how changes in solvent salinity affect the binding of proteins to DNA.
