Cell Differentiation during Sexual Development of the Fungus Sordaria macrospora Requires ATP Citrate Lyase Activity
Cell Differentiation during Sexual Development of the Fungus Sordaria macrospora Requires ATP Citrate Lyase Activity
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大孢子菌有性发育过程中的细胞分化需要 ATP 柠檬酸裂解酶活性
DOI：
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发表时间：
1999
影响因子：
5.3
通讯作者：
U. Kück
中科院分区：
文献类型：
--
作者：
M. Nowrousian;S. Masloff;S. Pöggeler;U. Kück
摘要在有性发育过程中，大多数丝状真菌的菌丝细胞分化为典型的子实体。在这里，我们描述了分离和表征的Sordaria macrospora发育突变体per 5，它表现出不育表型与缺陷的子实体成熟。细胞学研究表明，突变株只形成芽孢前体，没有任何成熟的孢子。使用索引的粘粒库，我们能够通过DNA介导的转化来补充突变体的育性。一个单一的粘粒克隆，携带一个3.5 kb的区域能够补充突变表型，已被确定。对3.5 kb区域的测序显示了一个2.1 kb的开放阅读框架，被一个66 bp的内含子中断。预测的多肽（674个氨基酸）与真核生物ATP柠檬酸裂解酶（ATP citrate lyases，acl）具有62 ~ 65%的同源性，并将该基因命名为acl 1。S.大孢霉ACL 1多肽的分子量为73 kDa，如通过用血凝素（HA）表位标记的ACL 1多肽进行的蛋白质印迹分析所证实的。HA标记的多肽的免疫原位检测表明，ACL位于胞质溶胶内。突变acl 1基因的测序揭示了编码区内的1个核苷酸的转换，导致在预测的多肽内的氨基酸取代。进一步的证据表明，ACL 1是必要的子实体成熟来自实验中，截断和突变版本的acl 1基因用于转化。这些拷贝都不能在转化的per 5受体菌株中重建可育表型。辅酶A通常参与细胞溶质乙酰辅酶A（乙酰辅酶A）的形成，其用于脂肪酸和甾醇的生物合成。与从野生型菌株获得的值相比，突变菌株的蛋白质提取物显示酶活性急剧降低。ACL表达的时间过程的调查表明，ACL是专门诱导在性周期的开始，并产生乙酰辅酶A，这很可能是一个先决条件，子实体形成在性发育的后期阶段。我们讨论了ACL活性对S.大孢子虫。
ABSTRACT During sexual development, mycelial cells from most filamentous fungi differentiate into typical fruiting bodies. Here, we describe the isolation and characterization of the Sordaria macrosporadevelopmental mutant per5, which exhibits a sterile phenotype with defects in fruiting body maturation. Cytological investigations revealed that the mutant strain forms only ascus precursors without any mature spores. Using an indexed cosmid library, we were able to complement the mutant to fertility by DNA-mediated transformation. A single cosmid clone, carrying a 3.5-kb region able to complement the mutant phenotype, has been identified. Sequencing of the 3.5-kb region revealed an open reading frame of 2.1 kb interrupted by a 66-bp intron. The predicted polypeptide (674 amino acids) shows significant homology to eukaryotic ATP citrate lyases (ACLs), with 62 to 65% amino acid identity, and the gene was named acl1. The molecular mass of the S. macrospora ACL1 polypeptide is 73 kDa, as was verified by Western blot analysis with a hemagglutinin (HA) epitope-tagged ACL1 polypeptide. Immunological in situ detection of the HA-tagged polypeptide demonstrated that ACL is located within the cytosol. Sequencing of the mutant acl1 gene revealed a 1-nucleotide transition within the coding region, resulting in an amino acid substitution within the predicted polypeptide. Further evidence that ACL1 is essential for fruiting body maturation comes from experiments in which truncated and mutated versions of theacl1 gene were used for transformation. None of these copies was able to reconstitute the fertile phenotype in transformed per5 recipient strains. ACLs are usually involved in the formation of cytosolic acetyl coenzyme A (acetyl-CoA), which is used for the biosynthesis of fatty acids and sterols. Protein extracts from the mutant strain showed a drastic reduction in enzymatic activity compared to values obtained from the wild-type strain. Investigation of the time course of ACL expression suggests that ACL is specifically induced at the beginning of the sexual cycle and produces acetyl-CoA, which most probably is a prerequisite for fruiting body formation during later stages of sexual development. We discuss the contribution of ACL activity to the life cycle of S. macrospora.
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影响因子：
1.5
作者：
M. Goodrich-Tanrikulu;Kelly Howe;A. Stafford;M. A. Nelson
通讯作者：
M. Goodrich-Tanrikulu;Kelly Howe;A. Stafford;M. A. Nelson
ATP-柠檬酸裂解酶和磷酸酶抑制剂 2 上被多功能蛋白激酶（糖原合酶激酶 3 样激酶）磷酸化的位点序列。
影响因子：
2.9
作者：
Ramakrishna,S;D'Angelo,G;Benjamin,WB
通讯作者：
Benjamin,WB
胰岛素对 ATP-柠檬酸裂解酶磷酸化的影响：肽 A 和肽 B 磷酸化的调节。
影响因子：
2.9
作者：
Ramakrishna,S;Murthy,KS;Benjamin,WB
通讯作者：
Benjamin,WB
DOI：
10.1016/s0021-9258(19)52474-7
发表时间：
1981-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. Pierce;J. Palmer;H. Keutmann;J. Avruch
通讯作者：
M. Pierce;J. Palmer;H. Keutmann;J. Avruch
粗糙脉孢菌性发育所需的假定鼠李糖半乳糖醛酸酶。
影响因子：
3.3
作者：
Nelson,MA;Merino,ST;Metzenberg,RL
通讯作者：
Metzenberg,RL