Heme-Peroxidase 2 Modulated by POU2F1 and SOX5 is Involved in Pigmentation in Pacific Oyster (Crassostrea gigas)
Heme-Peroxidase 2 Modulated by POU2F1 and SOX5 is Involved in Pigmentation in Pacific Oyster (Crassostrea gigas)
复制标题
DOI：
10.1007/s10126-022-10098-z
复制
发表时间：
2022-03
影响因子：
3
通讯作者：
Yue Min;Qi Li;Hong Yu
中科院分区：
文献类型：
--
作者：
Yue Min;Qi Li;Hong Yu
在软体类贝类中经常观察到颜色多态性，而贝壳色素沉着的分子调控尚不清楚。过氧化物酶是参与黑色素形成的关键酶。在这里，我们鉴定了一个血红素过氧化物酶2基因（CgHPX2），并表征了CgHPX2在太平洋牡蛎中的表达模式和转录调控。组织表达分析表明，cghpx2是一个地幔特异性基因，与白壳牡蛎相比，cghpx2主要表达在黑壳牡蛎的边缘地幔中。原位杂交表明，cghpx2在黑壳彩色牡蛎的外地幔褶皱内外表面都检测到强信号，而白壳牡蛎的阳性信号主要集中在外地幔褶皱的外表面。在胚胎和幼虫中，cghpx2在黑、白壳色牡蛎的trochophore阶段均有高表达。cghpx2在时间上的定位主要在棘球蚴和钙化壳幼虫的壳腺和边缘地幔中检测到。此外，克隆了cghpx2 5 '侧链2227 bp的序列，该序列包含一个假定的核心启动子区域和许多潜在的转录因子结合位点。进一步的荧光素酶分析实验证实，POU结构域、2类、转录因子1 （POU2F1）和SRY-box转录因子5 （SOX5）通过结合cghpx2基因的特定启动子区域参与了cghpx2基因的转录调控。经过cgpou2f1和cgsox5rna干扰后，ghpx2基因表达明显降低。这些结果提示，cghp2f1和cgsox5可能是两个重要的转录因子，正调控cghpx2基因的表达，提高了我们对软体动物壳色素沉着转录调控的认识。
Color polymorphism is frequently observed in molluscan shellfish, while the molecular regulation of shell pigmentation is not well understood. Peroxidase is a key enzyme involved in melanogenesis. Here, we identified a heme-peroxidase 2 gene (CgHPX2), and characterized the expression patterns and transcriptional regulation ofCgHPX2in the Pacific oysterCrassostrea gigas. Tissues expression analysis showed thatCgHPX2was a mantle-specific gene and primarily expressed in the edge mantle in black shell color oyster compared with white shell oyster. In situ hybridization showed that strong signals forCgHPX2were detected in the both inner and outer surface of the outer fold of mantle in the black shell color oyster, whereas positive signals in white shell oyster were mainly localized in the outer surface of the outer fold of mantle. In the embryos and larvae, a high expression level ofCgHPX2was detected in the trochophore stage in both black and white shell color oysters. The temporal localization ofCgHPX2was mainly detected in the shell gland and edge mantle of trochophore and calcified shell larvae, respectively. In addition, a 2227 bp of 5′ flanking region sequence ofCgHPX2was cloned, which contained a presumed core promoter region and many potential transcription factor binding sites. Further luciferase assay experiment confirmed that POU domain, class 2, transcription factor 1 (POU2F1), and SRY-box transcription factor 5 (SOX5) were involved in transcriptional regulation ofCgHPX2gene through binding to its specific promoter region. AfterCgPOU2F1andCgSOX5RNA interference, theCgHPX2gene expression was significantly decreased. These results suggested thatCgPOU2F1andCgSOX5might be two important transcription factors that positively regulated the expression ofCgHPX2gene, improving our understanding of the transcriptional regulation of molluscan shell pigmentation.