P1B-type ATPase ion pumps that transport heavy metal ions across cellular membranes are essential for plant growth and development. To date, a genomic comparison overview of the family in rice, maize and sorghum is no...P1B-type ATPase ion pumps that transport heavy metal ions across cellular membranes are essential for plant growth and development. To date, a genomic comparison overview of the family in rice, maize and sorghum is not yet available. In this study, a total of 31 heavy metal P1B-type ATPase （HMA） genes were identified, including 9 in rice, 11 each from maize and sorghum. They were classified into two distinct subfamilies based on their sequence composition and phylogenetic relationship. Four pairs of HMA genes were expanded via gene duplication with tandemly duplicated. Comprehensive analyses were performed to investigate the expression profiles of HMA genes in various tissues by using quantitative real-time PCR. Some HMA members exhibited abundant and tissue-specific expression patterns. Moreover, most of the genes were found to be differentially expressed under the Cu/Cd treatment. This study will facilitate further studies on P1B-type ATPase family and provide valuable hints for the functional validation in rice, maize and sorghum.展开更多
The regulatory mechanisms of cytoplasmic Ca2＋ after myocardial infarction-induced Ca2＋ overload involve secretory pathway Ca2＋-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi ...The regulatory mechanisms of cytoplasmic Ca2＋ after myocardial infarction-induced Ca2＋ overload involve secretory pathway Ca2＋-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi apparatus on Ca2＋ overload after cerebral ischemia and reperfusion remains unclear. Four-vessel occlusion rats were used as animal models of cerebral ischemia. The expression of secretory pathway Ca2＋-ATPase 1 in the cortex and hippocampus was detected by immunoblotting, and Ca2＋ concentrations in the cytoplasm and Golgi vesicles were determined. Results showed an overload of cytoplasmic Ca2＋ during ischemia and reperfusion that reached a peak after reperfusion. Levels of Golgi Ca2＋ showed an opposite effect. The expression of Golgi-specific secretory pathway Ca2＋-ATPase 1 in the cortex and hippocampus decreased before ischemia and reperfusion, and increased after reperfusion for 6 hours. This variation was similar to the alteration of calcium in separated Golgi vesicles. These results indicate that the Golgi apparatus participates in the formation and alleviation of calcium overload, and that secretory pathway Ca2＋-ATPase 1 tightly responds to ischemia and reperfusion in nerve cells. Thus, we concluded that secretory pathway Ca2＋-ATPase 1 plays an essential role in cytosolic calcium regulation and its expression can be used as a marker of Golgi stress, responding to cerebral ischemia and reperfusion. The secretory pathway Ca2＋-ATPase 1 can be an important neuroprotective target of ischemic stroke.展开更多
P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phy...P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phylogenetic tree analysis showed that OsHMA6 belonged to the Cu/Ag subgroup of the HMA family and had a close evolutionary relationship with OsHMA9.Amino acid sequence alignment showed 82.78%consistency between OsHMA6 and OsHMA9.OsHMA6 expressed in all organs at different growth stages,including spikelet,and abundant in leaf blades,however,OsHMA9 most strongly expressed in roots,but very low in spikelet.Excessive Cu^2+can up-regulate the expression of OsHMA6 and OsHMA9 in rice seedlings.The heterologous expression in yeast showed that OsHMA6 can significantly rescue the growth of yeast strain CM52 when supplied with 3 or 6 mmol/L Cu^2+.Compared with the empty vector pYES2,the Cu concentration in OsHMA6-pYES2 decreased by 23.4%and 30.3%under 3 or 6 mmol/L Cu2+,respectively.Subcellular localization revealed that OsHMA6 was located in the plasma membrane.These results suggested that OsHMA6,similar to OsHMA9,is likely a copper efflux protein located in the plasma membrane.展开更多
基金supported by the Special Fund for Agro-Scientific Research in the Public Interest of China(Grant No.201403015)
文摘P1B-type ATPase ion pumps that transport heavy metal ions across cellular membranes are essential for plant growth and development. To date, a genomic comparison overview of the family in rice, maize and sorghum is not yet available. In this study, a total of 31 heavy metal P1B-type ATPase （HMA） genes were identified, including 9 in rice, 11 each from maize and sorghum. They were classified into two distinct subfamilies based on their sequence composition and phylogenetic relationship. Four pairs of HMA genes were expanded via gene duplication with tandemly duplicated. Comprehensive analyses were performed to investigate the expression profiles of HMA genes in various tissues by using quantitative real-time PCR. Some HMA members exhibited abundant and tissue-specific expression patterns. Moreover, most of the genes were found to be differentially expressed under the Cu/Cd treatment. This study will facilitate further studies on P1B-type ATPase family and provide valuable hints for the functional validation in rice, maize and sorghum.
基金supported by the National Natural Science Foundation of China,No.81171239
文摘The regulatory mechanisms of cytoplasmic Ca2＋ after myocardial infarction-induced Ca2＋ overload involve secretory pathway Ca2＋-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi apparatus on Ca2＋ overload after cerebral ischemia and reperfusion remains unclear. Four-vessel occlusion rats were used as animal models of cerebral ischemia. The expression of secretory pathway Ca2＋-ATPase 1 in the cortex and hippocampus was detected by immunoblotting, and Ca2＋ concentrations in the cytoplasm and Golgi vesicles were determined. Results showed an overload of cytoplasmic Ca2＋ during ischemia and reperfusion that reached a peak after reperfusion. Levels of Golgi Ca2＋ showed an opposite effect. The expression of Golgi-specific secretory pathway Ca2＋-ATPase 1 in the cortex and hippocampus decreased before ischemia and reperfusion, and increased after reperfusion for 6 hours. This variation was similar to the alteration of calcium in separated Golgi vesicles. These results indicate that the Golgi apparatus participates in the formation and alleviation of calcium overload, and that secretory pathway Ca2＋-ATPase 1 tightly responds to ischemia and reperfusion in nerve cells. Thus, we concluded that secretory pathway Ca2＋-ATPase 1 plays an essential role in cytosolic calcium regulation and its expression can be used as a marker of Golgi stress, responding to cerebral ischemia and reperfusion. The secretory pathway Ca2＋-ATPase 1 can be an important neuroprotective target of ischemic stroke.
基金the Agricultural Science and Technology Innovation Program Cooperation and Innovation Mission(Grant No.CAAS-XTCX2016001)Shenzhen Science and Technology Projects(Grant No.JSGG20160608160725473)+1 种基金China Postdoctoral Science Foundation(Grant No.2018M641558)Fundamental Research Funds for Science,Technology and Innovation Commission of Shenzhen Municipality(Grant No.JCYJ20160530191619099).
文摘P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phylogenetic tree analysis showed that OsHMA6 belonged to the Cu/Ag subgroup of the HMA family and had a close evolutionary relationship with OsHMA9.Amino acid sequence alignment showed 82.78%consistency between OsHMA6 and OsHMA9.OsHMA6 expressed in all organs at different growth stages,including spikelet,and abundant in leaf blades,however,OsHMA9 most strongly expressed in roots,but very low in spikelet.Excessive Cu^2+can up-regulate the expression of OsHMA6 and OsHMA9 in rice seedlings.The heterologous expression in yeast showed that OsHMA6 can significantly rescue the growth of yeast strain CM52 when supplied with 3 or 6 mmol/L Cu^2+.Compared with the empty vector pYES2,the Cu concentration in OsHMA6-pYES2 decreased by 23.4%and 30.3%under 3 or 6 mmol/L Cu2+,respectively.Subcellular localization revealed that OsHMA6 was located in the plasma membrane.These results suggested that OsHMA6,similar to OsHMA9,is likely a copper efflux protein located in the plasma membrane.
