Characterization of Hydra Type IV Collagen

doi:10.1074/jbc.M005871200

Characterization of Hydra Type IV Collagen

TYPE IV COLLAGEN IS ESSENTIAL FOR HEAD REGENERATION AND ITS EXPRESSION IS UP-REGULATED UPON EXPOSURE TO GLUCOSE*

From the Wellcome Trust Centre for Cell-Matrix Research, School of Biological Sciences, University of Manchester, Manchester M13 9PT, United Kingdom, the ^§Department of Anatomy and Cell Biology, University of Kansas Medical Centre, Kansas City, Kansas 66160, the ^‖Institut fur Biochemie I, University of Regensburg, Regensburg, D-8400, Germany, and the ^¶Department of Biomedical Sciences, Southwest Missouri State University, Springfield, Missouri 65804-0094

Abstract

Hydra vulgaris mesoglea is a primitive basement membrane that also exhibits some features of an interstitial matrix. We have characterized cDNAs that encode the full-length hydra α1(IV) chain. The 5169-base pair transcript encodes a protein of 1723 amino acids, including an interrupted 1455-residue collagenous domain and a 228-residue C-terminal noncollagenous domain. N-terminal sequence analyses of collagen IV peptides suggest the molecule is homotrimeric. Denatured hydra type IV collagen protein occurs as dimers and higher order aggregates held together by nonreducible cross-links. Hydra collagen IV exhibits no functional evidence for the presence of a 7 S domain. Type IV collagen is expressed by the ectoderm along the entire longitudinal axis of the animal but is most intense at the base of the tentacles at the site of battery cell transdifferentiation. Antisense studies show that inhibition of collagen IV translation causes a blockage in head regeneration, indicating its importance in normal hydra development. Exposure of adult hydra to 15 mm glucose resulted in up-regulation of type IV collagen mRNA levels within 48 h and significant thickening of the mesoglea within 14 days, suggesting that basement membrane thickening seen in diabetes may be, in evolutionary terms, an ancient glucose-mediated response.

Footnotes

↵* This work was funded by grants from the Dr. Hadwen Trust for Humane Research and the Royal Society (to R. B.-H.), National Institutes of Health Grant DK47840 (to M. P. S.), and Grant SFB 521/A4 from the Deutsche Forschungsgemeinschaft (to R. D.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) .
↵** To whom correspondence should be addressed: Wellcome Trust Centre for Cell-Matrix Research, 2.205 Stopford, School of Biological Sciences, University of Manchester, Manchester M13 9PT, UK. Fax: 161-275-5082; E-mail“ Ray.Boot-Handford@man.ac.uk.
Published, JBC Papers in Press, August 23, 2000, DOI 10.1074/jbc.M005871200
↵2 M. Sarras, Jr., unpublished observations.
Abbreviations:

ECM

extracellular matrix

BM

basement membrane

HM

hydra medium

LEP

localized electroporation

UTR

untranslated region

bp

base pair(s)

kb

kilobase(s)

CHAPS

3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid

FITC

fluorescein isothiocyanate

contig

group of overlapping clones
- Received July 5, 2000.
- Revision received August 14, 2000.
The American Society for Biochemistry and Molecular Biology, Inc.