M142.2 (cut-6), a novel Caenorhabditis elegans matrix gene important for dauer body shape.
The cuticle of the nematode Caenorhabditis elegans is a collagenous extracellular matrix which forms the exoskeleton and defines the shape of the worm. We have characterized the C. elegans gene M142.2, and we show that this is a developmentally regulated gene important for cuticle structure. Transgenic worms expressing M142.2 promoter fused to green fluorescent protein showed that M142.2 is expressed in late embryos and L2d predauers, in the hypodermal cells which synthesize the cuticle. The same temporal pattern was seen by RT-PCR using RNA purified from specific developmental stages. A recombinant fragment of M142.2 was expressed in Escherichia coli and used to raise an antiserum. Immunohistochemistry using the antiserum localized M142.2 to the periphery of the alae of L1 and dauers, forming two longitudinal ribbons over the hypodermal cells. Loss-of-function of M142.2 by RNAi resulted in a novel phenotype: dumpy dauers which lacked alae. M142.2 therefore plays a major role in the assembly of the alae and the morphology of the dauer cuticle; because of its similarity to the other cut genes of the cuticle, we have named the gene cut-6.
Muriel
Joaquin M
JM
Department of Cell and Molecular Biology, Northwestern University Medical School, 303 E Chicago Ave, Chicago, IL 60611, USA.
Brannan
Mike
M
Taylor
Kathryn
K
Johnstone
Iain L
IL
Lithgow
Gordon J
GJ
Tuckwell
Danny
D
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D019521:Body Patterning / Q000235:genetics*; D017173:Caenorhabditis elegans / Q000033:anatomy & histology / Q000196:embryology / Q000502:physiology*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D004625:Embryo, Nonmammalian; D004926:Escherichia coli / Q000235:genetics; D005109:Extracellular Matrix / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D007106:Immune Sera; D007814:Larva / Q000502:physiology*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D040521:Subcutaneous Tissue / Q000196:embryology / Q000502:physiology
D029742:Caenorhabditis elegans Proteins; C477644:Cut-6 protein, C elegans; D007106:Immune Sera; D011994:Recombinant Proteins