Molecular characterization of a RNA polymerase (RNAP) II (DNA directed) polypeptide H (POLR2H) in Pacific white shrimp (Litopenaeus vannamei) and its role in response to high-pH stress.
Molecular characterization of a RNA polymerase (RNAP) II (DNA directed) polypeptide H (POLR2H) in Pacific white shrimp (Litopenaeus vannamei) and its role in response to high-pH stress.
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DOI：
10.1016/j.fsi.2019.12.012
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发表时间：
2019-12
影响因子：
4.7
通讯作者：
Zhi-Zhan Luo;Hui Sun;Jing-Wen Guo;Peng Luo;Chao Hu;Wen Huang;H. Shu
中科院分区：
文献类型：
--
作者：
Zhi-Zhan Luo;Hui Sun;Jing-Wen Guo;Peng Luo;Chao Hu;Wen Huang;H. Shu
RNA聚合酶（RNA - polymerase， RNAP） II (DNA-directed) （POLR2）基因对于细胞在环境胁迫下的生存能力以及生物信息从DNA到RNA的传递至关重要。然而，polr2基因在甲壳类动物中的功能和特征尚不清楚。本研究从凡纳滨对虾（Litopenaeus vannamei）中分离到aPOLR2HcDNA，并将其命名为asLv-POLR2H。全长772 bp，包含一个32 bp的5 ' -未翻译区（UTR）、一个284 bp的3 ' -非翻译区（UTR）和一个456 bp的开放阅读框（ORF），编码一个含有151个氨基酸的lv - polr2h蛋白，分子量为17.21 kDa。v- polr2h蛋白仅含有一个功能域，不含跨膜结构域，主要位于细胞核内。theLv-POLR2HmRNA的表达在所有选择的组织中普遍存在，在鳃中表达水平最高。原位杂交（ISH）分析表明，lv - polr2h主要分布在次生鳃丝中，在pH、低盐度和高浓度NO2−和NH4+的胁迫下，lv - polr2h在鳃中的转录水平受到显著影响，表明lv - polr2hin在各种物理胁迫下可能在鳃组织中发挥作用。具体而言，在高ph胁迫下，flv - polr2hvia siRNA的敲低显著降低了对虾的存活率，表明其在高ph胁迫响应中发挥了关键作用。本研究为polr2hin对虾在高pH胁迫下的作用提供了第一个证据，并为甲壳类动物对高pH胁迫的分子调控提供了新的见解。
RNA polymerase (RNAP) II (DNA-directed) (POLR2) genes are essential for cell viability under environmental stress and for the transfer of biological information from DNA to RNA. However, the function and characteristics ofPOLR2genes in crustaceans are still unknown. In the present study, aPOLR2HcDNA was isolated from Pacific white shrimp (Litopenaeus vannamei) and designated asLv-POLR2H. The full-lengthLv-POLR2HcDNA is 772 bp in length and contains a 32-bp 5′- untranslated region (UTR), a 284-bp 3′- UTR with a poly (A) sequence, and an open reading frame (ORF) of 456 bp encoding anLv-POLR2Hprotein of 151 amino acids with a deduced molecular weight of 17.21 kDa. TheLv-POLR2Hprotein only contains one functional domain, harbors no transmembrane domains and mainly locates in the nucleus. The expression of theLv-POLR2HmRNA was ubiquitously detected in all selected tissues, with the highest level in the gills. In situ hybridization (ISH) analysis showed thatLv-POLR2Hwas mainly located in the secondary gill filaments, the transcript levels ofLv-POLR2Hin the gills were found to be significantly affected after challenge by pH, low salinity and high concentrations of NO2−and NH4+, indicating thatLv-POLR2Hin gill tissues might play roles under various physical stresses. Specifically, under high-pH stress, knockdown ofLv-POLR2Hvia siRNA significantly decreased the survival rate of the shrimp, indicating its key roles in the response to high-pH stress. Our study may provide the first evidence of the role ofPOLR2Hin shrimp responding to high-pH stress and provides new insight into molecular regulation in response to high pH in crustaceans.