Development of metazoans requires the production of multiple cell types and the morphogenesis of complex multicellular structures. Genetic studies have established roles for glycosylation and proteoglycan biosynthesis in developmental signaling. The nematode Caenorhabditis elegans is a good model system to study the role of glycoconjugates in multicellular organisms as well as a prototypic model for parasitic nematodes because of the knowledge of its complete genomic sequence and development. In this R2 I (Exploratory / Developmental) grant application, we wish to explore a novel potential functional linkage between expression of glycoconjugates and developmental events in C. elegans. Srf-9, srf-8 and srf-4 (surface) are single loci mutants that have multiple defects, including uncoordinated movement, protruding vulva, abnormal egg laying, and defective copulatory bursae and gonad morphology, yet they were isolated based on their ectopic surface binding to a lectin which binds to N-acetylglucosamine. Mutations in srf-9, srf-8 and srf-4 interact with mutations in the lin-12 gene, which a member of the LIN- 1 2/NOTCH family of receptor proteins that mediate cell-cell interactions to specify cell fate during development. We will test the hypothesis that the srf-9, srf-8 and srf-4 genes are involved in the biosynthesis, post-translational processing or secretion of glycoconjugates, and therefore link developmental events with glycoconjugates expression. Specifically, the proposed work will: 1) Define the glycoconjugates affected in srf-9, srf-8 and srf-4 mutants, through the structural characterization of cuticle glycan fractions, 2) Identify the srf-9, srf-8 and srf-4 genes through molecular cloning using a combination of Single Nucleotide Polymorphism (SNP) mapping, and marker rescue experiments. The information derived from these studies will provide the tools to study the role of these genes in developmental events in the nematode C. elegans. Because complex phenotypes are the hallmark of several human genetic disorders, including some that affect glycosylation, the proposed studies will establish the feasibility of using C. elegans srf mutants as a model for glycosylation disorders in humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21GM066156-02
Application #
6647188
Study Section
Pathobiochemistry Study Section (PBC)
Program Officer
Marino, Pamela
Project Start
2002-08-15
Project End
2004-07-31
Budget Start
2003-08-01
Budget End
2004-07-31
Support Year
2
Fiscal Year
2003
Total Cost
$121,125
Indirect Cost
Name
Boston University
Department
Biochemistry
Type
Schools of Dentistry
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02118
Burket, Christopher T; Higgins, Christina E; Hull, Lynn C et al. (2006) The C. elegans gene dig-1 encodes a giant member of the immunoglobulin superfamily that promotes fasciculation of neuronal processes. Dev Biol 299:193-205
Berninsone, Patricia M; Hirschberg, Carlos B (2002) The nematode Caenorhabditis elegans as a model to study the roles of proteoglycans. Glycoconj J 19:325-30