In eukaryotic organisms,the most common internal modification of messenger RNA(m RNA)is N6-methyladenosine(m6A).This modification can be dynamically and reversibly controlled by specific enzymes known as m6A writers a...In eukaryotic organisms,the most common internal modification of messenger RNA(m RNA)is N6-methyladenosine(m6A).This modification can be dynamically and reversibly controlled by specific enzymes known as m6A writers and erasers.The fat-mass and obesity-associated protein(FTO)catalyzes RNA demethylation and plays a critical role in various physiological and pathological processes.Our research identified dynamic alterations in both m6A and FTO during the assembly of primordial follicles,with an inverse relationship observed for m6A levels and nuclear-localized FTO expression.Application of Fto small interfering RNA(si RNA)altered the expression of genes related to cell proliferation,hormone regulation,and cell chemotaxis,and affected RNA alternative splicing.Overexpression of the full-length Fto gene led to changes in m6A levels,alternative splicing of Cdk5,cell proliferation,cell cycle progression,and proportion of primordial follicles.Conversely,overexpression of Fto lacking a nuclear localization signal(NLS)did not significantly alter m6A levels or primordial follicle assembly.These findings suggest that FTO,localized in the nucleus but not in the cytoplasm,regulates RNA m6A demethylation and plays a role in cell proliferation,cell cycle progression,and primordial follicle assembly.These results highlight the potential of m6A and its eraser FTO as possible biomarkers and therapeutic targets.展开更多
为了探究绵羊FTO(Fat mass and obesity-associated protein)基因的结构和功能,试验采用绵羊皮肤组织RNA-seq获得的CDS序列和生物信息学方法对FTO基因的核苷酸序列的相似性、氨基酸序列、蛋白理化性质、蛋白二、三级结构、亚细胞定位和...为了探究绵羊FTO(Fat mass and obesity-associated protein)基因的结构和功能,试验采用绵羊皮肤组织RNA-seq获得的CDS序列和生物信息学方法对FTO基因的核苷酸序列的相似性、氨基酸序列、蛋白理化性质、蛋白二、三级结构、亚细胞定位和互作蛋白进行研究。结果表明,绵羊FTO基因CDS区为1518 bp,共编码505个氨基酸,其中亮氨酸(Leu)含量最多(11.1%),组氨酸(His)含量最少(2.6%);FTO蛋白分子式C_(2609)H_(4053)N_(709)O_(771)S_(27),分子的质量为58553.79,理论等电点(pI)为5.28,半衰期为30h,结构不稳定,属于亲水性蛋白,不具有信号肽和跨膜结构域;FTO主要分布在细胞外基质中;多肽链中含有43.17%α螺旋结构、38.61%无规则卷曲、11.49%延伸链和6.73%β折叠结构;FTO与FOXO1、CREB1、ALKBH8、ALKBH5、YTHDF1、YTHDF3、YTHDF2、METTL14、WTAP、METTL310个蛋白可能存在互作。说明绵羊的FTO蛋白主要与RNA甲基化调控相关蛋白互作,为进一步研究其功能提供参考。展开更多
基金supported by the Natural Science Foundation of Shandong Province,China (ZR2017MC033)National Key Research and Development Program of China (2023YFD1300504)Taishan Scholar Construction Foundation of Shandong Province,China (ts20190946)。
文摘In eukaryotic organisms,the most common internal modification of messenger RNA(m RNA)is N6-methyladenosine(m6A).This modification can be dynamically and reversibly controlled by specific enzymes known as m6A writers and erasers.The fat-mass and obesity-associated protein(FTO)catalyzes RNA demethylation and plays a critical role in various physiological and pathological processes.Our research identified dynamic alterations in both m6A and FTO during the assembly of primordial follicles,with an inverse relationship observed for m6A levels and nuclear-localized FTO expression.Application of Fto small interfering RNA(si RNA)altered the expression of genes related to cell proliferation,hormone regulation,and cell chemotaxis,and affected RNA alternative splicing.Overexpression of the full-length Fto gene led to changes in m6A levels,alternative splicing of Cdk5,cell proliferation,cell cycle progression,and proportion of primordial follicles.Conversely,overexpression of Fto lacking a nuclear localization signal(NLS)did not significantly alter m6A levels or primordial follicle assembly.These findings suggest that FTO,localized in the nucleus but not in the cytoplasm,regulates RNA m6A demethylation and plays a role in cell proliferation,cell cycle progression,and primordial follicle assembly.These results highlight the potential of m6A and its eraser FTO as possible biomarkers and therapeutic targets.
文摘为了探究绵羊FTO(Fat mass and obesity-associated protein)基因的结构和功能,试验采用绵羊皮肤组织RNA-seq获得的CDS序列和生物信息学方法对FTO基因的核苷酸序列的相似性、氨基酸序列、蛋白理化性质、蛋白二、三级结构、亚细胞定位和互作蛋白进行研究。结果表明,绵羊FTO基因CDS区为1518 bp,共编码505个氨基酸,其中亮氨酸(Leu)含量最多(11.1%),组氨酸(His)含量最少(2.6%);FTO蛋白分子式C_(2609)H_(4053)N_(709)O_(771)S_(27),分子的质量为58553.79,理论等电点(pI)为5.28,半衰期为30h,结构不稳定,属于亲水性蛋白,不具有信号肽和跨膜结构域;FTO主要分布在细胞外基质中;多肽链中含有43.17%α螺旋结构、38.61%无规则卷曲、11.49%延伸链和6.73%β折叠结构;FTO与FOXO1、CREB1、ALKBH8、ALKBH5、YTHDF1、YTHDF3、YTHDF2、METTL14、WTAP、METTL310个蛋白可能存在互作。说明绵羊的FTO蛋白主要与RNA甲基化调控相关蛋白互作,为进一步研究其功能提供参考。
