The AP-3 Clathrin-associated Complex Is Essential for Embryonic and Larval Development in Caenorhabditis elegans

  • Shim, Jaegal (Department of Biology, Yonsei University) ;
  • Lee, Junho (Department of Biology, Yonsei University)
  • Received : 2005.02.21
  • Accepted : 2005.03.03
  • Published : 2005.06.30

Abstract

The adaptor protein (AP) complexes are involved in membrane transport of many proteins. There are 3 AP complexes in C. elegans unlike mammals that have four. To study the biological functions of the AP-3 complexes of C. elegans, we sought homologues of the mouse and human genes that encode subunits of the AP-3 complexes by screening C. elegans genomic and EST sequences. We identified single copies of homologues of the ${\mu}3$, ${\sigma}3$, ${\beta}3$ and ${\delta}$ genes. The medium chain of AP-3 is encoded by a single gene in C. elegans but two different genes in mammals. Since there are no known mutations in these genes in C. elegans, we performed RNAi to assess their functions in development. RNAi of each of the genes caused embryonic and larval lethal phenotypes. APM-3 is expressed in most cells, particularly strongly in spermatheca and vulva. We conclude that the products of the C. elegans ${\mu}3$, ${\sigma}3$, ${\beta}3$ and d genes are essential for embryogenesis and larval development.

Keywords

Acknowledgement

Supported by : Ministry of Science and Technology

References

  1. Brenner, S. (1974) The genetics of Caenorhabditis elegans. Genetics 77, 71-94
  2. Dell'Angelica, E. C., Shotelersuk, V., Aguilar, R. C., Gahl, W. A., and Bonifacino, J. S. (1999) Altered trafficking of lysosomal proteins in Hermansky-Pudlak syndrome due to mutations in the beta 3A subunit of the AP-3 adaptor. Mol. Cell 3, 11-21 https://doi.org/10.1016/S1097-2765(00)80170-7
  3. Dell'Angelica, E. C., Mullins, C., Caplan, S., and Bonifacino, J. S. (2000) Lysosome-related organelles. FASEB J. 14, 1265- 1278 https://doi.org/10.1096/fj.14.10.1265
  4. Feng, L., Seymour, A. B., Jiang, S., To, A., Peden, A. A., et al. (1999) The beta3A subunit gene (Ap3b1) of the AP-3 adaptor complex is altered in the mouse hypopigmentation mutant pearl, a model for Hermansky-Pudlak syndrome and night blindness. Hum. Mol. Genet. 8, 323-330 https://doi.org/10.1093/hmg/8.2.323
  5. Fraser, A. G., Kamath, R. S., Zipperlen, P., Martinez-Campos, M., Sohrmann, M., et al. (2000) Functional genomic analysis of C. elegans chromosome I by systematic RNA interference. Nature 408, 325-330 https://doi.org/10.1038/35042517
  6. Kantheti, P., Qiao, X., Diaz, M. E., Peden, A. A., Meyer, G. E., et al. (1998) Mutation in AP-3 delta in the mocha mouse links endosomal transport to storage deficiency in platelets, melanosomes, and synaptic vesicles. Neuron 21, 111-122 https://doi.org/10.1016/S0896-6273(00)80519-X
  7. Koushika, S. P. and Nonet, M. L. (2000) Sorting and transport in C. elegans: a model system with a sequenced genome. Curr. Opin. Cell Biol. 12, 517-523 https://doi.org/10.1016/S0955-0674(00)00125-3
  8. Kretzschmar, D., Poeck, B., Roth, H., Ernst, R., Keller, A., et al. (2000) Defective pigment granule biogenesis and aberrant behavior caused by mutations in the Drosophila AP-3beta adaptin gene ruby. Genetics 155, 213-223
  9. Le Borgne, R., Alconada, A., Bauer, U., and Hoflack, B. (1998) The mammalian AP-3 adaptor-like complex mediates the in tracellular transport of lysosomal membrane glycoproteins. J. Biol. Chem. 273, 29451-29461 https://doi.org/10.1074/jbc.273.45.29451
  10. Lee, J., Jongeward, G. D., and Sternberg, P. W. (1994) unc-101, a gene required for many aspects of Caenorhabditis elegans development and behavior, encodes a clathrin-associated protein. Genes Dev. 8, 60-73 https://doi.org/10.1101/gad.8.1.60
  11. Lloyd, V., Sinclair, D. A., Wennberg, R., Warner, T. S., Honda, B. M., et al. (1999) A genetic and molecular characterization of the garnet gene of Drosophila melanogaster. Genome 42, 1183-1193 https://doi.org/10.1139/gen-42-6-1183
  12. Lloyd, V., Ramaswami, M., and Kramer, H. (1998) Not just pretty eyes: Drosophila eye-colour mutations and lysosomal delivery. Trends Cell Biol. 8, 257-259 https://doi.org/10.1016/S0962-8924(98)01270-7
  13. Maniatis, T., Fritsch, E. F., and Sambrook, J. (1989) Molecular Cloning, Cold Spring Harber Laboratory Press, Cold Spring Harber, NY
  14. Meyer, C., Zizioli, D., Lausmann, S., Eskelinen, E. L., Hamann, J., et al. (2000) mu1A-adaptin-deficient mice: lethality, loss of AP-1 binding and rerouting of mannose 6-phosphate receptors. EMBO J. 19, 2193-2203 https://doi.org/10.1093/emboj/19.10.2193
  15. Mullins, C., Hartnell, L. M., and Bonifacino, J. S. (2000) Distinct requirements for the AP-3 adaptor complex in pigment granule and synaptic vesicle biogenesis in Drosophila melanogaster. Mol. Gen. Genet. 263, 1003-1014 https://doi.org/10.1007/PL00008688
  16. Newman, L. S., McKeever, M. O., Okano, H. J., and Darnell, R. B. (1995) Beta-NAP, a cerebellar degeneration antigen, is a neuron-specific vesicle coat protein. Cell 82, 773-783 https://doi.org/10.1016/0092-8674(95)90474-3
  17. Odorizzi, G., Cowles, C. R., and Emr, S. D. (1998) The AP-3 complex: a coat of many colours. Trends Cell Biol. 8, 282-288 https://doi.org/10.1016/S0962-8924(98)01295-1
  18. Ooi, C. E., Moreira, J. E., Dell'Angelica, E. C., Poy, G., Wassarman, D. A., et al. (1997) Altered expression of a novel adaptin leads to defective pigment granule biogenesis in the Drosophila eye color mutant garnet. EMBO J. 16, 4508- 4518 https://doi.org/10.1093/emboj/16.15.4508
  19. Pevsner, J., Volknandt, W., Wong, B. R., and Scheller, R. H. (1994) Two rat homologs of clathrin-associated adaptor proteins. Gene 146, 279-283 https://doi.org/10.1016/0378-1119(94)90306-9
  20. Robinson, M. S. and Bonifacino, J. S. (2001) Adaptor-related proteins. Curr. Opin. Cell Biol. 13, 444-453 https://doi.org/10.1016/S0955-0674(00)00235-0
  21. Shim, J. and Lee, J. (2000) Molecular genetic analysis of apm-2 and aps-2, genes encoding the medium and small chains of the AP-2 clathrin-associated protein complex in the nematode Caenorhabditis elegans. Mol. Cells 10, 309-316
  22. Shim, J., Sternberg, P. W., and Lee, J. (2000) Distinct and redundant functions of mu1 medium chains of the AP-1 clathrin-associated protein complex in the nematode Caenorhabditis elegans. Mol. Biol. Cell. 11, 2743-2756
  23. Simpson, F., Bright, N. A., West, M. A., Newman, L. S., Darnell, R. B., et al. (1996) A novel adaptor-related protein complex. J. Cell Biol. 133, 749-760 https://doi.org/10.1083/jcb.133.4.749
  24. Simpson, F., Peden, A. A., Christopoulou, L., and Robinson, M. S. (1997) Characterization of the adaptor-related protein complex, AP-3. J. Cell Biol. 137, 835-845 https://doi.org/10.1083/jcb.137.4.835
  25. Stepp, J. D., Huang, K., and Lemmon, S. K. (1997) The yeast adaptor protein complex, AP-3, is essential for the efficient delivery of alkaline phosphatase by the alternate pathway to the vacuole. J. Cell Biol. 139, 1761-1774 https://doi.org/10.1083/jcb.139.7.1761
  26. Zizioli, D., Meyer, C., Guhde, G., Saftig, P., von Figura, K., et al. (1999) Early embryonic death of mice deficient in gammaadaptin. J. Biol. Chem. 274, 5385-5390 https://doi.org/10.1074/jbc.274.9.5385