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. 1997 Jul 16;16(14):4194–4204. doi: 10.1093/emboj/16.14.4194

Suppressors of YCK-encoded yeast casein kinase 1 deficiency define the four subunits of a novel clathrin AP-like complex.

H R Panek 1, J D Stepp 1, H M Engle 1, K M Marks 1, P K Tan 1, S K Lemmon 1, L C Robinson 1
PMCID: PMC1170045  PMID: 9250663

Abstract

In Saccharomyces cerevisiae, the redundant YCK1 and YCK2 genes (Yeast Casein Kinase 1) are required for viability. We describe here the molecular analysis of four mutations that eliminate the requirement for Yck activity. These mutations alter proteins that resemble the four subunits of clathrin adaptors (APs), with highest sequence similarity to those of the recently identified AP-3 complex. The four yeast subunits are associated in a high-molecular-weight complex. These proteins have no essential function and are not redundant for function with other yeast AP-related proteins. Combination of suppressor mutations with a clathrin heavy chain mutation (chc1-ts) confers no synthetic growth defects. However, a yck(ts) mutation shows a strong synthetic growth defect with chc1-ts. Moreover, endocytosis of Ste3p is dramatically decreased in yck(ts) cells and is partially restored by the AP suppressor mutations. These results suggest that vesicle trafficking at the plasma membrane requires the activity of Yck protein kinases, and that the new AP-related complex may participate in this process.

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Selected References

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  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Bar-Zvi D., Mosley S. T., Branton D. In vivo phosphorylation of clathrin-coated vesicle proteins from rat reticulocytes. J Biol Chem. 1988 Mar 25;263(9):4408–4415. [PubMed] [Google Scholar]
  3. Brockman J. L., Gross S. D., Sussman M. R., Anderson R. A. Cell cycle-dependent localization of casein kinase I to mitotic spindles. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9454–9458. doi: 10.1073/pnas.89.20.9454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cukierman E., Huber I., Rotman M., Cassel D. The ARF1 GTPase-activating protein: zinc finger motif and Golgi complex localization. Science. 1995 Dec 22;270(5244):1999–2002. doi: 10.1126/science.270.5244.1999. [DOI] [PubMed] [Google Scholar]
  5. Davis N. G., Horecka J. L., Sprague G. F., Jr Cis- and trans-acting functions required for endocytosis of the yeast pheromone receptors. J Cell Biol. 1993 Jul;122(1):53–65. doi: 10.1083/jcb.122.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fish K. J., Cegielska A., Getman M. E., Landes G. M., Virshup D. M. Isolation and characterization of human casein kinase I epsilon (CKI), a novel member of the CKI gene family. J Biol Chem. 1995 Jun 23;270(25):14875–14883. doi: 10.1074/jbc.270.25.14875. [DOI] [PubMed] [Google Scholar]
  7. Graves P. R., Haas D. W., Hagedorn C. H., DePaoli-Roach A. A., Roach P. J. Molecular cloning, expression, and characterization of a 49-kilodalton casein kinase I isoform from rat testis. J Biol Chem. 1993 Mar 25;268(9):6394–6401. [PubMed] [Google Scholar]
  8. Hicke L., Riezman H. Ubiquitination of a yeast plasma membrane receptor signals its ligand-stimulated endocytosis. Cell. 1996 Jan 26;84(2):277–287. doi: 10.1016/s0092-8674(00)80982-4. [DOI] [PubMed] [Google Scholar]
  9. Hoekstra M. F., Liskay R. M., Ou A. C., DeMaggio A. J., Burbee D. G., Heffron F. HRR25, a putative protein kinase from budding yeast: association with repair of damaged DNA. Science. 1991 Aug 30;253(5023):1031–1034. doi: 10.1126/science.1887218. [DOI] [PubMed] [Google Scholar]
  10. Hoffman C. S., Winston F. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987;57(2-3):267–272. doi: 10.1016/0378-1119(87)90131-4. [DOI] [PubMed] [Google Scholar]
  11. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kirchhausen T., Davis A. C., Frucht S., Greco B. O., Payne G. S., Tubb B. AP17 and AP19, the mammalian small chains of the clathrin-associated protein complexes show homology to Yap17p, their putative homolog in yeast. J Biol Chem. 1991 Jun 15;266(17):11153–11157. [PubMed] [Google Scholar]
  13. Kirchhausen T. Identification of a putative yeast homolog of the mammalian beta chains of the clathrin-associated protein complexes. Mol Cell Biol. 1990 Nov;10(11):6089–6090. doi: 10.1128/mcb.10.11.6089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nakayama Y., Goebl M., O'Brine Greco B., Lemmon S., Pingchang Chow E., Kirchhausen T. The medium chains of the mammalian clathrin-associated proteins have a homolog in yeast. Eur J Biochem. 1991 Dec 5;202(2):569–574. doi: 10.1111/j.1432-1033.1991.tb16409.x. [DOI] [PubMed] [Google Scholar]
  15. Newman L. S., McKeever M. O., Okano H. J., Darnell R. B. Beta-NAP, a cerebellar degeneration antigen, is a neuron-specific vesicle coat protein. Cell. 1995 Sep 8;82(5):773–783. doi: 10.1016/0092-8674(95)90474-3. [DOI] [PubMed] [Google Scholar]
  16. Page L. J., Robinson M. S. Targeting signals and subunit interactions in coated vesicle adaptor complexes. J Cell Biol. 1995 Nov;131(3):619–630. doi: 10.1083/jcb.131.3.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pauloin A., Thurieau C. The 50 kDa protein subunit of assembly polypeptide (AP) AP-2 adaptor from clathrin-coated vesicles is phosphorylated on threonine-156 by AP-1 and a soluble AP50 kinase which co-purifies with the assembly polypeptides. Biochem J. 1993 Dec 1;296(Pt 2):409–415. doi: 10.1042/bj2960409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Phan H. L., Finlay J. A., Chu D. S., Tan P. K., Kirchhausen T., Payne G. S. The Saccharomyces cerevisiae APS1 gene encodes a homolog of the small subunit of the mammalian clathrin AP-1 complex: evidence for functional interaction with clathrin at the Golgi complex. EMBO J. 1994 Apr 1;13(7):1706–1717. doi: 10.1002/j.1460-2075.1994.tb06435.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roach P. J. Control of glycogen synthase by hierarchal protein phosphorylation. FASEB J. 1990 Sep;4(12):2961–2968. [PubMed] [Google Scholar]
  20. Robinson L. C., Menold M. M., Garrett S., Culbertson M. R. Casein kinase I-like protein kinases encoded by YCK1 and YCK2 are required for yeast morphogenesis. Mol Cell Biol. 1993 May;13(5):2870–2881. doi: 10.1128/mcb.13.5.2870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Roth A. F., Davis N. G. Ubiquitination of the yeast a-factor receptor. J Cell Biol. 1996 Aug;134(3):661–674. doi: 10.1083/jcb.134.3.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rowles J., Slaughter C., Moomaw C., Hsu J., Cobb M. H. Purification of casein kinase I and isolation of cDNAs encoding multiple casein kinase I-like enzymes. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9548–9552. doi: 10.1073/pnas.88.21.9548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Seaman M. N., Sowerby P. J., Robinson M. S. Cytosolic and membrane-associated proteins involved in the recruitment of AP-1 adaptors onto the trans-Golgi network. J Biol Chem. 1996 Oct 11;271(41):25446–25451. doi: 10.1074/jbc.271.41.25446. [DOI] [PubMed] [Google Scholar]
  24. Seeger M., Payne G. S. Selective and immediate effects of clathrin heavy chain mutations on Golgi membrane protein retention in Saccharomyces cerevisiae. J Cell Biol. 1992 Aug;118(3):531–540. doi: 10.1083/jcb.118.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Simpson F., Bright N. A., West M. A., Newman L. S., Darnell R. B., Robinson M. S. A novel adaptor-related protein complex. J Cell Biol. 1996 May;133(4):749–760. doi: 10.1083/jcb.133.4.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Simpson F., Peden A. A., Christopoulou L., Robinson M. S. Characterization of the adaptor-related protein complex, AP-3. J Cell Biol. 1997 May 19;137(4):835–845. doi: 10.1083/jcb.137.4.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stamnes M. A., Rothman J. E. The binding of AP-1 clathrin adaptor particles to Golgi membranes requires ADP-ribosylation factor, a small GTP-binding protein. Cell. 1993 Jun 4;73(5):999–1005. doi: 10.1016/0092-8674(93)90277-w. [DOI] [PubMed] [Google Scholar]
  28. Stepp J. D., Pellicena-Palle A., Hamilton S., Kirchhausen T., Lemmon S. K. A late Golgi sorting function for Saccharomyces cerevisiae Apm1p, but not for Apm2p, a second yeast clathrin AP medium chain-related protein. Mol Biol Cell. 1995 Jan;6(1):41–58. doi: 10.1091/mbc.6.1.41. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sullivan M. A., Cannon J. F., Webb F. H., Bock R. M. Antisuppressor mutation in Escherichia coli defective in biosynthesis of 5-methylaminomethyl-2-thiouridine. J Bacteriol. 1985 Jan;161(1):368–376. doi: 10.1128/jb.161.1.368-376.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Traub L. M., Ostrom J. A., Kornfeld S. Biochemical dissection of AP-1 recruitment onto Golgi membranes. J Cell Biol. 1993 Nov;123(3):561–573. doi: 10.1083/jcb.123.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tuazon P. T., Traugh J. A. Casein kinase I and II--multipotential serine protein kinases: structure, function, and regulation. Adv Second Messenger Phosphoprotein Res. 1991;23:123–164. [PubMed] [Google Scholar]
  32. Vancura A., O'Connor A., Patterson S. D., Mirza U., Chait B. T., Kuret J. Isolation and properties of YCK2, a Saccharomyces cerevisiae homolog of casein kinase-1. Arch Biochem Biophys. 1993 Aug 15;305(1):47–53. doi: 10.1006/abbi.1993.1391. [DOI] [PubMed] [Google Scholar]
  33. Vancura A., Sessler A., Leichus B., Kuret J. A prenylation motif is required for plasma membrane localization and biochemical function of casein kinase I in budding yeast. J Biol Chem. 1994 Jul 29;269(30):19271–19278. [PubMed] [Google Scholar]
  34. Wang P. C., Vancura A., Mitcheson T. G., Kuret J. Two genes in Saccharomyces cerevisiae encode a membrane-bound form of casein kinase-1. Mol Biol Cell. 1992 Mar;3(3):275–286. doi: 10.1091/mbc.3.3.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wang X., Hoekstra M. F., DeMaggio A. J., Dhillon N., Vancura A., Kuret J., Johnston G. C., Singer R. A. Prenylated isoforms of yeast casein kinase I, including the novel Yck3p, suppress the gcs1 blockage of cell proliferation from stationary phase. Mol Cell Biol. 1996 Oct;16(10):5375–5385. doi: 10.1128/mcb.16.10.5375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Watanabe T. K., Shimizu F., Nagata M., Takaichi A., Fujiwara T., Nakamura Y., Takahashi E., Hirai Y. Cloning, expression pattern and mapping to 12p 13.2 --> p13.1 of CLAPS3, a gene encoding a novel clathrin-adaptor small chain. Cytogenet Cell Genet. 1996;73(3):214–217. doi: 10.1159/000134341. [DOI] [PubMed] [Google Scholar]
  37. Wilde A., Brodsky F. M. In vivo phosphorylation of adaptors regulates their interaction with clathrin. J Cell Biol. 1996 Nov;135(3):635–645. doi: 10.1083/jcb.135.3.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zanolari B., Raths S., Singer-Krüger B., Riezman H. Yeast pheromone receptor endocytosis and hyperphosphorylation are independent of G protein-mediated signal transduction. Cell. 1992 Nov 27;71(5):755–763. doi: 10.1016/0092-8674(92)90552-n. [DOI] [PubMed] [Google Scholar]
  39. Zhai L., Graves P. R., Robinson L. C., Italiano M., Culbertson M. R., Rowles J., Cobb M. H., DePaoli-Roach A. A., Roach P. J. Casein kinase I gamma subfamily. Molecular cloning, expression, and characterization of three mammalian isoforms and complementation of defects in the Saccharomyces cerevisiae YCK genes. J Biol Chem. 1995 May 26;270(21):12717–12724. doi: 10.1074/jbc.270.21.12717. [DOI] [PubMed] [Google Scholar]
  40. Ziman M., Chuang J. S., Schekman R. W. Chs1p and Chs3p, two proteins involved in chitin synthesis, populate a compartment of the Saccharomyces cerevisiae endocytic pathway. Mol Biol Cell. 1996 Dec;7(12):1909–1919. doi: 10.1091/mbc.7.12.1909. [DOI] [PMC free article] [PubMed] [Google Scholar]

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