Cloning and expression of a type IX-like collagen in tissues of the ascidian Ciona intestinalis

Collagens are highly preserved proteins in invertebrates and vertebrates. To identify the collagens in urochordates, the total RNAextracted from the pharynx of the ascidian Ciona intestinalis was hybridized with a heterologous probe specific for the echinodermParacentrotus lividus fibrillar type I-like larval collagen. Using this probe, two main bands (i.e. 6 and 2.8 kb mRNA) were observed onNorthern blot hybridization. The cDNA library prepared from poly(A) +RNA extracted from pharyngeal tissue was screened and a cDNAthat specifies a type IX-like collagen was identified. This molecule presents a conceptual open reading frame for a protein containing 734amino acids. In particular, we showed a 1a chain type IX-like collagen characterized by three short triple-helical domains interspersed withfour non-triple-helical sequences, with structural features of fibril-associated collagens with interrupted triple-helices (FACIT) collagens.Northern blot hybridizations indicate a 2.8 kb transcript size. Sequence comparison indicated homology (47.64%, 48.95%) between the typeIX-like collagen of C. intestinalis and mouse and human type IX collagen. In situ hybridization of tunic and pharynx tissues shows thepresence of transcripts in connective tissue cells.