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. 1991 Dec 1;115(5):1237–1247. doi: 10.1083/jcb.115.5.1237

Altered expression of an L1-specific, O-linked cuticle surface glycoprotein in mutants of the nematode Caenorhabditis elegans

PMCID: PMC2289243  PMID: 1955471

Abstract

Mouse mAb M38 was used in indirect immunofluorescence experiments to detect a stage-specific antigen on the surface of the first larval stage (L1) of the free-living nematode Caenorhabditis elegans, and to detect alterations in the apparent expression of this antigen in two distinct classes of C. elegans mutants. In previously described srf-2 and srf-3 mutants (Politz S. M., M. T. Philipp, M. Estevez, P.J. O'Brien, and K. J. Chin. 1990. Proc. Natl. Acad. Sci. USA. 87:2901- 2905), the antigen is not detected on the surface of any stage. Conversely, in srf-(yj43) and other similar mutants, the antigen is expressed on the surface of the first through the fourth (L4) larval stages. To understand the molecular basis of these alterations, the antigen was characterized in gel immunoblotting experiments. After SDS- PAGE separation and transfer to nitrocellulose, M38 detected a protein antigen in extracts of wild-type L1 populations. The antigen was sensitive to digestion by Pronase and O-glycanase (endo-alpha-N- acetylgalactosaminidase), suggesting that it is an O-linked glycoprotein. This antigen was not detected in corresponding extracts of wild-type L4s or srf-2 or srf-3 L1s, but was detected in extracts of srf-(yj43) L4s. The antigen-defective phenotype of srf-3 was epistatic to the heterochronic mutant phenotype of srf-(yj43) in immunofluorescence tests of the srf-3 srf-(yj43) double mutant, suggesting that srf-(yj43) causes incorrect regulation of a pathway of antigen formation that requires wild-type srf-3 activity.

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Selected References

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  1. Ambros V. A hierarchy of regulatory genes controls a larva-to-adult developmental switch in C. elegans. Cell. 1989 Apr 7;57(1):49–57. doi: 10.1016/0092-8674(89)90171-2. [DOI] [PubMed] [Google Scholar]
  2. Ambros V., Horvitz H. R. Heterochronic mutants of the nematode Caenorhabditis elegans. Science. 1984 Oct 26;226(4673):409–416. doi: 10.1126/science.6494891. [DOI] [PubMed] [Google Scholar]
  3. Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  5. Cox G. N., Hirsh D. Stage-specific patterns of collagen gene expression during development of Caenorhabditis elegans. Mol Cell Biol. 1985 Feb;5(2):363–372. doi: 10.1128/mcb.5.2.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cox G. N., Kusch M., Edgar R. S. Cuticle of Caenorhabditis elegans: its isolation and partial characterization. J Cell Biol. 1981 Jul;90(1):7–17. doi: 10.1083/jcb.90.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cox G. N., Staprans S., Edgar R. S. The cuticle of Caenorhabditis elegans. II. Stage-specific changes in ultrastructure and protein composition during postembryonic development. Dev Biol. 1981 Sep;86(2):456–470. doi: 10.1016/0012-1606(81)90204-9. [DOI] [PubMed] [Google Scholar]
  8. Deshaies R. J., Kepes F., Böhni P. C. Genetic dissection of the early stages of protein secretion in yeast. Trends Genet. 1989 Mar;5(3):87–93. doi: 10.1016/0168-9525(89)90032-2. [DOI] [PubMed] [Google Scholar]
  9. Emmons S. W., Klass M. R., Hirsh D. Analysis of the constancy of DNA sequences during development and evolution of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1333–1337. doi: 10.1073/pnas.76.3.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Grencis R. K., Crawford C., Pritchard D. I., Behnke J. M., Wakelin D. Immunization of mice with surface antigens from the muscle larvae of Trichinella spiralis. Parasite Immunol. 1986 Nov;8(6):587–596. doi: 10.1111/j.1365-3024.1986.tb00872.x. [DOI] [PubMed] [Google Scholar]
  11. Ikegaki N., Kennett R. H. Glutaraldehyde fixation of the primary antibody-antigen complex on nitrocellulose paper increases the overall sensitivity of immunoblot assay. J Immunol Methods. 1989 Nov 30;124(2):205–210. doi: 10.1016/0022-1759(89)90354-2. [DOI] [PubMed] [Google Scholar]
  12. Kobata A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem. 1979 Nov 15;100(1):1–14. doi: 10.1016/0003-2697(79)90102-7. [DOI] [PubMed] [Google Scholar]
  13. Kramer J. M., Cox G. N., Hirsh D. Expression of the Caenorhabditis elegans collagen genes col-1 and col-2 is developmentally regulated. J Biol Chem. 1985 Feb 10;260(3):1945–1951. [PubMed] [Google Scholar]
  14. Kramer J. M., Johnson J. J., Edgar R. S., Basch C., Roberts S. The sqt-1 gene of C. elegans encodes a collagen critical for organismal morphogenesis. Cell. 1988 Nov 18;55(4):555–565. doi: 10.1016/0092-8674(88)90214-0. [DOI] [PubMed] [Google Scholar]
  15. Kusch M., Edgar R. S. Genetic studies of unusual loci that affect body shape of the nematode Caenorhabditis elegans and may code for cuticle structural proteins. Genetics. 1986 Jul;113(3):621–639. doi: 10.1093/genetics/113.3.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Maizels R. M., de Savigny D., Ogilvie B. M. Characterization of surface and excretory-secretory antigens of Toxocara canis infective larvae. Parasite Immunol. 1984 Jan;6(1):23–37. doi: 10.1111/j.1365-3024.1984.tb00779.x. [DOI] [PubMed] [Google Scholar]
  19. Philipp M., Davis T. B., Storey N., Carlow C. K. Immunity in filariasis: perspectives for vaccine development. Annu Rev Microbiol. 1988;42:685–716. doi: 10.1146/annurev.mi.42.100188.003345. [DOI] [PubMed] [Google Scholar]
  20. Philipp M., Parkhouse R. M., Ogilvie B. M. Changing proteins on the surface of a parasitic nematode. Nature. 1980 Oct 9;287(5782):538–540. doi: 10.1038/287538a0. [DOI] [PubMed] [Google Scholar]
  21. Philipp M., Rumjaneck F. D. Antigenic and dynamic properties of helminth surface structures. Mol Biochem Parasitol. 1984 Mar;10(3):245–268. doi: 10.1016/0166-6851(84)90025-2. [DOI] [PubMed] [Google Scholar]
  22. Politz J. C., Edgar R. S. Overlapping stage-specific sets of numerous small collagenous polypeptides are translated in vitro from Caenorhabditis elegans RNA. Cell. 1984 Jul;37(3):853–860. doi: 10.1016/0092-8674(84)90420-3. [DOI] [PubMed] [Google Scholar]
  23. Politz S. M., Chin K. J., Herman D. L. Genetic analysis of adult-specific surface antigenic differences between varieties of the nematode Caenorhabditis elegans. Genetics. 1987 Nov;117(3):467–476. doi: 10.1093/genetics/117.3.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Politz S. M., Philipp M., Estevez M., O'Brien P. J., Chin K. J. Genes that can be mutated to unmask hidden antigenic determinants in the cuticle of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1990 Apr;87(8):2901–2905. doi: 10.1073/pnas.87.8.2901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Smith H. V., Quinn R., Kusel J. R., Girdwood R. W. The effect of temperature and antimetabolites on antibody binding to the outer surface of second stage Toxocara canis larvae. Mol Biochem Parasitol. 1981 Dec;4(3-4):183–193. doi: 10.1016/0166-6851(81)90017-7. [DOI] [PubMed] [Google Scholar]
  26. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. von Mende N., Bird D. M., Albert P. S., Riddle D. L. dpy-13: a nematode collagen gene that affects body shape. Cell. 1988 Nov 18;55(4):567–576. doi: 10.1016/0092-8674(88)90215-2. [DOI] [PubMed] [Google Scholar]

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