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 RNA extracted from the pharynx of the ascidian Ciona intestinalis was hybridized with a heterologous probe specific for the echinoderm Paracentrotus lividus fibrillar type I-like larval collagen. Using this probe, two main bands (i.e. 6 and 2.8 kb mRNA) were observed on Northern blot hybridization. The cDNA library prepared from poly(A)+RNA extracted from pharyngeal tissue was screened and a cDNA that specifies a type IX-like collagen was identified. This molecule presents a conceptual open reading frame for a protein containing 734 amino acids. In particular, we showed a 1 alpha chain type IX-like collagen characterized by three short triple-helical domains interspersed with four 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 type IX-like collagen of C. intestinalis and mouse and human type IX collagen. In situ hybridization of tunic and pharynx tissues shows the presence of transcripts in connective tissue cells.
Vizzini
Aiti
A
Department of Animal Biology, University of Palermo, via Archirafi 18, 90123 Palermo, Italy.
Arizza
Vincenzo
V
Cervello
Melchiorre
M
Cammarata
Matteo
M
Gambino
Roberto
R
Parrinello
Nicolò
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002938:Ciona intestinalis / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D024081:Collagen Type IX / Q000096:biosynthesis* / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014158:Transcription, Genetic
D024081:Collagen Type IX; D012333:RNA, Messenger