Skip to main content
Log in

Novel aspects of the regulation of a cDNA (Arf1) from Chlamydomonas with high sequence identity to animal ADP-ribosylation factor 1

  • Research Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

ADP-ribosylation factor (ARF) is a highly conserved, low molecular mass (ca. 21 kDa) GTP-binding protein that has been implicated in vesicle trafficking and signal transduction in yeast and mammalian cells. However, little is known of ARF in plant systems. A putative ARF polypeptide was identifed in subcellular fractions of the green alga Chlamydomonas reinhardtii, based on [32P]GTP binding and immunoblot assays. A cDNA clone was isolated from Chlamydomonas (Arf1), which encodes a 20.7 kDa protein with 90% identity to human ARF1. Northern blot analyses showed that levels of Arf1 mRNA are highly regulated during 12 h/12 h light/dark (LD) cycles. A biphasic pattern of expression was observed: a transient peak of Arf1 mRNA occurred at the onset of the light period, which was followed ca. 12 h later by a more prominent peak in the early to mid-dark period. When LD-synchronized cells were shifted to continuous darkness, the dark-specific peak of Arf1 mRNA persisted, indicative of a circadian rhythm. The increase in Arf1 mRNA at the beginning of the light period, however, was shown to be light-dependent, and, moreover, dependent on photosynthesis, since it was prevented by DCMU. We conclude that the biphasic pattern of Arf1 mRNA accumulation during LD cycles is due to regulation by two different factors, light (which requires photosynthesis) and the circadian clock. Thus, these studies identify a novel pattern of expression for a GTP-binding protein gene.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Amor JC, Harrison DH, Kahn RA, Ringe D: Structure of the human ADP-ribosylation factor 1 complexed with GDP. Nature 372: 704–708 (1994).

    PubMed  Google Scholar 

  2. Balch WE: Small GTP-binding proteins in vesicular transport. Trends Biochem Sci 15: 473–477 (1990).

    Article  PubMed  Google Scholar 

  3. Barr FA, Leyte A, Huttner WB: Trimeric G proteins and vesicle formation. Trends Cell Biol 2: 91–94 (1992).

    Article  PubMed  Google Scholar 

  4. Bennett MK, Scheller RH: The molecular machinery for secretion is conserved from yeast to neurons. Proc Natl Acad Sci USA 90: 2559–2563 (1993).

    PubMed  Google Scholar 

  5. Bobak DA, Nightingale MS, Murtaugh JJ, Price SR, Moss J, Vaughn M: Molecular cloning, characterization, and expression of human ADP-ribosylation factors: two guanine nucleotide-dependent activators of cholera toxin. Proc Natl Acad Sci USA 86: 6101–6105 (1989).

    PubMed  Google Scholar 

  6. Bourne HR, Sanders DA, McCormick F: The GTPase superfamily: conserved structure and molecular mechanism. Nature 349: 117–127 (1991).

    Article  PubMed  Google Scholar 

  7. Brown HA, Gutowski S, Moomaw CR, Slaughter C, Sternweis PC: ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity. Cell 75: 1137–1144 (1993).

    Article  PubMed  Google Scholar 

  8. Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995 (1984).

    PubMed  Google Scholar 

  9. Clark J, Moore L, Krasinskas A, Way J, Battey J, Tamkun J, Kahn RA: Selective amplification of additional members of the ADP-ribosylation factor (ARF) family: cloning of additional human and Drosophila ARF-like genes. Proc Natl Acad Sci USA 90: 8952–8956 (1993).

    PubMed  Google Scholar 

  10. Cockcroft S, Thomas GMH, Fensome A, Geny B, Cunningham E, Gout I, Hiles I, Totty NF, Truong O, Hsuan JJ: Phospholipase D: a downstream effector of ARF in granulocytes. Science 263: 523–526 (1994).

    PubMed  Google Scholar 

  11. Dascher C, Balch WE: Dominant inhibitory mutants of ARF1 block endoplasmic reticulum to Golgi transport and trigger disassembly of the Golgi apparatus. J Biol Chem 269: 1437–1448 (1994).

    PubMed  Google Scholar 

  12. Edmunds LN: Cellular and Molecular Bases of Biological Clocks. Springer-Verlag, New York (1988).

    Google Scholar 

  13. Elazar Z, Orci L, Ostermann J, Amherdt M, Tanigawa G, Rothman JE: ADP-ribosylation factor and coatomer couple fusion to vesicle budding. J Cell Biol 124: 415–425 (1994).

    Article  PubMed  Google Scholar 

  14. Fabry S, Nab N, Huber H, Palme K, Jaenicke L, Schmitt R: The yptV1 gene encodes a small G-protein in the green alga Volvox carteri: gene structure and properties of the gene product. Gene 118: 153–162 (1992).

    PubMed  Google Scholar 

  15. Fabry S, Jacobsen A, Huber H, Palme K, Schmitt R: Structure, expression, and phylogenetic relationships of a family of ypt genes encoding small G-proteins in the green alga Volvox carteri. Curr Genet 24: 229–240 (1993).

    PubMed  Google Scholar 

  16. Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).

    PubMed  Google Scholar 

  17. Gilman AG: G proteins: transducers of receptorgenerated signals. Annu Rev Biochem 56: 615–649 (1987).

    PubMed  Google Scholar 

  18. Hall A, Paterson HF, Adamson P, Ridley AJ: Cellular responses regulated by rho-related small GTP-binding proteins. Phil Trans R Soc Lond B 340: 267–271 (1993).

    Google Scholar 

  19. Harris EH: The Chlamydomonas Sourcebook: A Comprehensive Guide to Biology and Laboratory Use. Academic Press, San Diego (1989).

    Google Scholar 

  20. Herrin D, Michaels A: Gene expression during the cell cycle of Chlamydomonas reinhardtii. In: Stein G, Stein J (eds) Recombinant DNA and Cell Proliferation, pp. 87–106. Academic Press, Orlando (1984).

    Google Scholar 

  21. Herrin DL, Schmidt GW: Trans-splicing of transcripts for the chloroplast psaA1 gene. In vivo requirement for nuclear gene products. J Biol Chem 263: 14601–14604 (1988).

    PubMed  Google Scholar 

  22. Herrin DL, Schmidt GW: Rapid, reversible staining of northern blots prior to hybridization. Bio Techniques 6: 196–200 (1988).

    Google Scholar 

  23. Herrin DL and Worley T: A rapid procedure for the Isolation of chloroplast DNA from Chlamydomonas using the TL-100 ultracentrifuge. Plant Mol Biol Rep 8: 292–296 (1990).

    Google Scholar 

  24. Higo H, Kishimoto N, Saito A, Higo K-I: Molecular cloning and characterization of a cDNA encoding a small GTP-binding protein related to mammalian ADP-ribosylation factor from rice. Plant Sci 100: 41–49 (1994).

    Article  Google Scholar 

  25. Hwang S, Herrin DL: Control of lhc gene transcription by the circadian clock in chlamydomonas reinhardtii. Plant Mol Biol 26: 557–569 (1994).

    Article  PubMed  Google Scholar 

  26. Jacobshagen S, Johnson CH: Circadian rhythms of gene expression in Chlamydomonas reinhardtii: circadian cycling of mRNA abundances of cabII, and possibly of beta-tubulin and cytochrome c. Eur J Cell Biol 64: 142–152 (1994).

    PubMed  Google Scholar 

  27. Kahn RA, Gilman AG: Purification of a protein cofactor required for ADP-ribosylation of the stimulatory regulatory component of adenylate cyclase by cholera toxin. J Biol Chem 259: 6228–6234 (1984).

    PubMed  Google Scholar 

  28. Kahn RA, Goddard C, Newkirk M: Chemical and immunological characterization of the 21 kDa ADP-ribosylation factor (ARF) of adenylate cyclase. J Biol Chem 263: 8282–8287 (1988).

    PubMed  Google Scholar 

  29. Kindle KL: Expression of a gene for a light-harvesting chlorophyll a/b-binding protein in Chlamydomonas reinhardtii: effects of light and acetate. Plant Mol Biol 9: 547–563 (1987).

    Google Scholar 

  30. Lee F-JS, Stevens LA, Hall LM, Murtaugh JJ, Kao YL, Moss J, Vaughn M: Characterization of class II and class III ADP-ribosylation factor genes and proteins in Drosophila melanogaster. J Biol Chem 269: 21555–21560 (1994).

    PubMed  Google Scholar 

  31. Memon AR, Clark GB, Thompson GA Jr: Identification of an ARF type low molecular mass GTP-binding protein in pea (Pisum sativum). Biochem Biophys Res Commun 193: 809–813 (1993).

    Article  PubMed  Google Scholar 

  32. Memon AR, Herrin DL, Thompson GA Jr: Intracellular translocation of a 28 kDa GTP-binding protein during osmotic shock-induced cell volume regulation in Dunaliella salina. Biochim Biophys Acta 1179: 11–22 (1993).

    Article  PubMed  Google Scholar 

  33. Memon AR, kawazoe R, Zhang X, Herrin DL, Thompson GA: Low molecular mass GTP-binding proteins in Chlamydomonas reinhardtii wild type and a wall-less stram. Characterization and comparison with GTP-binding proteins of Dunaliella salina. Plant Physiol Biochem 33: 225–234 (1995).

    Google Scholar 

  34. Monaco L, Murtaugh JJ, Newman KB, Tsai S-C, Moss J, Vaughn M: Selective amplification of an mRNA and related pseudogene for a human ADP-ribosylation factor, a guanine nucleotide-dependent protein activator of cholera toxin. Proc Natl Acad Sci USA 87: 2206–2210 (1990).

    PubMed  Google Scholar 

  35. Moreland DE: Mechanism of action of herbicides. Annu Rev Plant Physiol 31: 597–638 (1980).

    Article  Google Scholar 

  36. Palmer DJ, Helm JB, Beckers CJM, Orci L, Rothman JE: Binding of coatomer to Golgi membranes requires ADP-ribosylation factor. J Biol Chem 26: 12083–12089 (1993).

    Google Scholar 

  37. Price SR, Welsh CF, haun SR, Stanley SJ, Moss J, Vaughan M: Effects of phospholipid and GTP on recombinant ADP-ribosylation factors (ARFs). Molecular basis for differences in requirements for activity of mammalian ARFs. J Biol Chem 267: 17766–17772 (1992).

    PubMed  Google Scholar 

  38. Pryer NK, Wuestehube LJ, Schekman R: Vesiclemediated protein sorting. Annu Rev Biochem 61: 471–516 (1992).

    Article  PubMed  Google Scholar 

  39. Regad F, Bardet C, Tremousaygue D, Moisan A, Lescure B, Axelos M: cDNA cloning and expression of an Arabidopsis GTP-binding protein of the ARF family. FEBS Lett 316: 133–136 (1993).

    Article  PubMed  Google Scholar 

  40. Rothman JE, Orci L: Molecular dissection of the secretory pathway. Nature 355: 409–415 (1992).

    Article  PubMed  Google Scholar 

  41. Schloss JA: A Chlamydomonas gene encodes a G protein β subunit-like polypeptide. Mol Gen Genet 221: 443–452 (1992).

    Google Scholar 

  42. Serafini T, Orci L, Amherdt M, Brunner M, Kahn RA, Rothman JE: ADP-ribosylation factor is a subunit of the coat of Golgi derived COP-coated vesicles: a novel role for a GTP-binding protein. Cell 67: 239–253 (1991).

    Article  PubMed  Google Scholar 

  43. Stearns TM, Hoyt MA, Botstein D, Kahn RA: ADP-ribosylation factor is an essential protein in yeast and is encoded by two genes. Mol Cell Biol 10: 6690–6699 (1990).

    PubMed  Google Scholar 

  44. Szopa J, Sikorski AF: ARF-protein antisense potato displays stable ADP-ribosylation of 40 kDa protein. J Plant Physiol 145: 383–386 (1995).

    Google Scholar 

  45. Takai Y, Kaibuchi K, Kikuchi A, Kawata M: Small GTP-binding proteins. Int Rev Cytol 133: 187–230 (1992).

    PubMed  Google Scholar 

  46. Tamkun JW, Kahn RA, Kissinger M, Brizuela BJ, Rulka C, Scott MP, Kennison JA: The arflike gene encodes an essential GTP-binding protein in Drosophila. Proc Natl Acad Sci USA 88: 3120–3124 (1991).

    PubMed  Google Scholar 

  47. Tsuchiya M, Price SR, Tsai S-C, Moss J, Vaughn M: Molecular identification of ADP-ribosylation factor mRNAs and their expression in mammalian cells. J Biol Chem 266: 2772–2777 (1991).

    PubMed  Google Scholar 

  48. Verwoert II GS, Brown A, Slabas AR, Stuitje AR: Z Zea mays GTP-binding protein of the ARF family complements an Escherichia coli mutant with a temperaturesensitive malonyl-coenzyme A:acyl carrier protein transacylase. Plant Mol Biol 27: 629–633 (1995).

    PubMed  Google Scholar 

  49. Walker MW, Bobak DA, Tsai S-C, Moss J, Vaughan M: GTP but not GDP analogues promote association of ADP-ribosylation factors, 20-kDa protein activators of cholera toxin, with phospholipids and PC-12 cell membranes. J Biol Chem 267: 3230–3235 (1992).

    PubMed  Google Scholar 

  50. Zhang C-J, Rosenwald AG, Willingham MC, Skuntz S, Clark J, Kahn RA: Expression of a dominant allele of human ARF1 inhibits membrane traffic in vivo. J Cell Biol 124: 289–300 (1994).

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Memon, A.R., Hwang, S., Deshpande, N. et al. Novel aspects of the regulation of a cDNA (Arf1) from Chlamydomonas with high sequence identity to animal ADP-ribosylation factor 1. Plant Mol Biol 29, 567–577 (1995). https://doi.org/10.1007/BF00020985

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00020985

Key words

Navigation