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© The Rockefeller University Press, 0021-9525/1997//1171 $5.00
The Journal of Cell Biology, Volume 137, Number 5, , 1997 1171-1183


Article

Type IV Collagen Is Detectable in Most, but Not All, Basement Membranes of Caenorhabditis elegans and Assembles on Tissues That Do Not Express It



Patricia L. Graham, Jeffrey J. Johnson, Shaoru Wang, Marion H. Sibley, Malini C. Gupta, and James M. Kramer

Northwestern University Medical School, Department of Cell and Molecular Biology, Chicago, Illinois 60611

Type IV collagen in Caenorhabditis elegans is produced by two essential genes, emb-9 and let-2, which encode {alpha}1- and {alpha}2-like chains, respectively. The distribution of EMB-9 and LET-2 chains has been characterized using chain-specific antisera. The chains colocalize, suggesting that they may function in a single heterotrimeric collagen molecule. Type IV collagen is detected in all basement membranes except those on the pseudocoelomic face of body wall muscle and on the regions of the hypodermis between body wall muscle quadrants, indicating that there are major structural differences between some basement membranes in C. elegans. Using lacZ/green fluorescent protein (GFP) reporter constructs, both type IV collagen genes were shown to be expressed in the same cells, primarily body wall muscles, and some somatic cells of the gonad. Although the pharynx and intestine are covered with basement membranes that contain type IV collagen, these tissues do not express either type IV collagen gene. Using an epitope-tagged emb-9 construct, we show that type IV collagen made in body wall muscle cells can assemble into the pharyngeal, intestinal, and gonadal basement membranes. Additionally, we show that expression of functional type IV collagen only in body wall muscle cells is sufficient for C. elegans to complete development and be partially fertile. Since type IV collagen secreted from muscle cells only assembles into some of the basement membranes that it has access to, there must be a mechanism regulating its assembly. We propose that interaction with a cell surface–associated molecule(s) is required to facilitate type IV collagen assembly.


1. Abbreviations used in this paper: dH2O, distilled water; GFP, green fluorescent protein; HA, hemagglutinin; NDS, normal donkey serum.

Address all correspondence to James M. Kramer, Northwestern University Medical School, Department of Cell and Molecular Biology, 303 E. Chicago Ave., Chicago, IL 60611. Tel.: (312) 503-7644. Fax: (312) 5032522 or 7912. E-mail: jkramer{at}nwu.edu

Marion H. Sibley's current address is New England Biolabs, Beverly, MA 01915.



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