Department for the Study of Territory and its Resources, University of Genova, Corso Europa 26, 16132, Genova, Italy.
Mar Biotechnol (NY). 2012 Jun;14(3):281-93. doi: 10.1007/s10126-011-9415-2. Epub 2011 Nov 10.
We report here the complete cDNA sequence of a nonfibrillar collagen (COLch) isolated from the marine sponge Chondrosia reniformis, Nardo 1847 using a PCR approach. COLch cDNA consists of 2,563 nucleotides and includes a 5' untranslated region (UTR) of 136 nucleotides, a 3' UTR of 198 nucleotides, and an open reading frame encoding for a protein of 743 amino acids with an estimated M (r) of 72.12 kDa. The phylogenetic analysis on the deduced amino acid sequence of C-terminal end shows that the isolated sequence belongs to the short-chain spongin-like collagen subfamily, a nonfibrillar group of invertebrate collagens similar to type IV collagen. In situ hybridization analysis shows higher expression of COLch mRNA in the cortical part than in the inner part of the sponge. Therefore, COLch seems to be involved in the formation of C. reniformis ectosome, where it could play a key role in the attachment to the rocky substrata and in the selective sediment incorporation typical of these organisms. qPCR analysis of COLch mRNA level, performed on C. reniformis tissue culture models (fragmorphs), also demonstrates that this matrix protein is directly involved in sponge healing processes and that soluble silicates positively regulate its expression. These findings confirm the essential role of silicon in the fibrogenesis process also in lower invertebrates, and they should give a tool for a sustainable production of marine collagen in sponge mariculture.
我们在这里报告了一种从海洋海绵 Chondrosia reniformis 中分离出的非纤维胶原(COLch)的完整 cDNA 序列,使用 PCR 方法。COLch cDNA 由 2563 个核苷酸组成,包括 136 个核苷酸的 5'非翻译区(UTR)、198 个核苷酸的 3'UTR 和一个开放阅读框,编码 743 个氨基酸的蛋白质,估计 M(r)为 72.12 kDa。C 末端推断氨基酸序列的系统发育分析表明,分离出的序列属于短链海绵样胶原亚家族,这是一种与 IV 型胶原相似的无纤维的无脊椎动物胶原。原位杂交分析显示 COLch mRNA 在海绵的皮质部分的表达高于内部部分。因此,COLch 似乎参与了 C. reniformis 外体的形成,在那里它可能在附着到岩石基质和这些生物特有的选择性沉积物掺入中发挥关键作用。COLch mRNA 水平的 qPCR 分析,在 C. reniformis 组织培养模型( fragmorphs)上进行,也表明这种基质蛋白直接参与海绵愈合过程,并且可溶性硅酸盐正向调节其表达。这些发现证实了硅在低等无脊椎动物纤维化过程中的重要作用,并且它们应该为海绵养殖中海洋胶原的可持续生产提供一种工具。