Mining and characterization of ubiquitin E3 ligases expressed in the mouse testis.
Mining and characterization of ubiquitin E3 ligases expressed in the mouse testis.
复制标题
DOI：
10.1186/1471-2164-13-495
复制
发表时间：
2012-09-19
期刊：
影响因子：
4.4
通讯作者：
Han C
中科院分区：
文献类型：
--
作者：
Hou X;Zhang W;Xiao Z;Gan H;Lin X;Liao S;Han C
泛素介导的蛋白质修饰和降解被认为在哺乳动物精子发生中起重要作用。已知哺乳动物如小鼠和人的泛素活化酶、缀合酶和连接酶（E3）的目录。然而，在精子发生过程中表达的E3的系统表征尚未进行。在本研究中，我们着手从小鼠基因组中挖掘E3，并根据微阵列数据和生化测定来表征其表达模式、亚细胞定位和酶活性。我们鉴定了398个属于RING、U-box和HECT亚家族的推定E3，发现大多数基因在小鼠和人类之间是保守的。我们发现其中73个基因在睾丸中高度表达或特异表达。我们选择了10个推定的E3基因来检测它们的mRNA表达模式，并选择了几个基因来研究它们的亚细胞定位和E3连接酶活性。RT-PCR结果表明，所选基因均在睾丸中表达。一些推定的E3定位在细胞质中，而另一些则在细胞质和细胞核中。此外，所有选定的蛋白质都具有酶活性，如体外和体内测定所示。我们已经确定了大量的推定E3在小鼠精子发生过程中表达。其中，显著部分在睾丸中高度或特异性表达。亚细胞定位和酶活性分析表明，这些E3可能在哺乳动物精子发生中执行不同的功能。我们的研究结果可以作为一个初步的指导，该领域的进一步功能分析。
Ubiquitin-mediated protein modification and degradation are believed to play important roles in mammalian spermatogenesis. The catalogues of ubiquitin activating enzymes, conjugating enzymes, and ligases (E3s) have been known for mammals such as mice and humans. However, a systematic characterization of E3s expressed during spermatogenesis has not been carried out. In present study, we set out to mine E3s from the mouse genome and to characterize their expression pattern, subcellular localization, and enzymatic activities based on microarray data and biochemical assays. We identified 398 putative E3s belonging to the RING, U-box, and HECT subfamilies and found that most genes were conserved between mice and humans. We discovered that 73 of them were highly or specifically expressed in the testes based on the microarray expression data. We selected 10 putative E3 genes to examine their mRNA expression pattern, and several genes to study their subcellular localization and E3 ligase activity. RT-PCR results showed that all the selected genes were predominately expressed in the testis. Some putative E3s were localized in the cytoplasm while others were in both the cytoplasm and the nucleus. Moreover, all the selected proteins were enzymatically active as demonstrated by in vitro and in vivo assays. We have identified a large number of putative E3s that are expressed during mouse spermatogenesis. Among these, a significant portion is highly or specifically expressed in the testis. Subcellular localization and enzymatic activity assays suggested that these E3s might execute diverse functions in mammalian spermatogenesis. Our results may serve as an initial guide to the field for further functional analysis.
登录
查看更多内容
影响因子：
3.7
作者：
Li W;Bengtson MH;Ulbrich A;Matsuda A;Reddy VA;Orth A;Chanda SK;Batalov S;Joazeiro CA
通讯作者：
Joazeiro CA
DOI：
10.1073/pnas.0910267107
发表时间：
2010-02-02
影响因子：
11.1
作者：
An, Jee Young;Kim, Eun-A.;Kwon, Yong Tae
通讯作者：
Kwon, Yong Tae
DOI：
10.1073/pnas.0709854105
发表时间：
2008-06-17
影响因子：
11.1
作者：
Johnston, Daniel S.;Wright, William W.;Jelinsky, Scott A.
通讯作者：
Jelinsky, Scott A.
影响因子：
2.7
作者：
Kopanja D;Roy N;Stoyanova T;Hess RA;Bagchi S;Raychaudhuri P
通讯作者：
Raychaudhuri P
影响因子：
2.1
作者：
Ogawa, Miyuki;Mizugishi, Kiyomi;Aruga, Jun
通讯作者：
Aruga, Jun