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. 1998 Aug;10(8):1295–1306. doi: 10.1105/tpc.10.8.1295

Maize endosperm ADP-glucose pyrophosphorylase SHRUNKEN2 and BRITTLE2 subunit interactions

TW Greene 1, LC Hannah 1
PMCID: PMC144067  PMID: 9707530

Abstract

ADP-glucose pyrophosphorylase (AGP) represents a key regulatory step in polysaccharide synthesis in organisms ranging from bacteria to plants. Higher plant AGPs are complex in nature and are heterotetramers consisting of two similar but distinct subunits. How the subunits are assembled into enzymatically active polymers is not yet understood. Here, we address this issue by using naturally occurring null mutants of the Shrunken2 (Sh2) and Brittle2 (Bt2) loci of maize as well as the yeast two-hybrid expression system. In the absence of the maize endosperm large AGP subunit (SH2), the BT2 subunit remains as a monomer in the developing endosperm. In contrast, the SH2 protein, in the absence of BT2, is found in a complex of 100 kD. A direct interaction between SH2 and BT2 was proven when they were both expressed in yeast. Several motifs are essential for SH2:BT2 interaction because truncations removing the N or C terminus of either subunit eliminate SH2:BT2 interactions. Analysis of subunit interaction mutants (sim) also identified motifs essential for protein interactions.

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Selected References

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  1. Ainsworth C., Hosein F., Tarvis M., Weir F., Burrell M., Devos K. M., Gale M. D. Adenosine diphosphate glucose pyrophosphorylase genes in wheat: differential expression and gene mapping. Planta. 1995;197(1):1–10. doi: 10.1007/BF00239933. [DOI] [PubMed] [Google Scholar]
  2. Ballicora M. A., Laughlin M. J., Fu Y., Okita T. W., Barry G. F., Preiss J. Adenosine 5'-diphosphate-glucose pyrophosphorylase from potato tuber. Significance of the N terminus of the small subunit for catalytic properties and heat stability. Plant Physiol. 1995 Sep;109(1):245–251. doi: 10.1104/pp.109.1.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bhave M. R., Lawrence S., Barton C., Hannah L. C. Identification and molecular characterization of shrunken-2 cDNA clones of maize. Plant Cell. 1990 Jun;2(6):581–588. doi: 10.1105/tpc.2.6.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Creech R G. Genetic Control of Carbohydrate Synthesis in Maize Endosperm. Genetics. 1965 Dec;52(6):1175–1186. doi: 10.1093/genetics/52.6.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dickinson D. B., Preiss J. Presence of ADP-Glucose Pyrophosphorylase in Shrunken-2 and Brittle-2 Mutants of Maize Endosperm. Plant Physiol. 1969 Jul;44(7):1058–1062. doi: 10.1104/pp.44.7.1058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fields S., Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989 Jul 20;340(6230):245–246. doi: 10.1038/340245a0. [DOI] [PubMed] [Google Scholar]
  8. Fields S., Sternglanz R. The two-hybrid system: an assay for protein-protein interactions. Trends Genet. 1994 Aug;10(8):286–292. doi: 10.1016/0168-9525(90)90012-u. [DOI] [PubMed] [Google Scholar]
  9. Giroux M. J., Hannah L. C. ADP-glucose pyrophosphorylase in shrunken-2 and brittle-2 mutants of maize. Mol Gen Genet. 1994 May 25;243(4):400–408. doi: 10.1007/BF00280470. [DOI] [PubMed] [Google Scholar]
  10. Giroux M. J., Shaw J., Barry G., Cobb B. G., Greene T., Okita T., Hannah L. C. A single mutation that increases maize seed weight. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5824–5829. doi: 10.1073/pnas.93.12.5824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Greene T. W., Chantler S. E., Kahn M. L., Barry G. F., Preiss J., Okita T. W. Mutagenesis of the potato ADPglucose pyrophosphorylase and characterization of an allosteric mutant defective in 3-phosphoglycerate activation. Proc Natl Acad Sci U S A. 1996 Feb 20;93(4):1509–1513. doi: 10.1073/pnas.93.4.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hannah L. C., Nelson O. E., Jr Characterization of ADP-glucose pyrophosphorylase from shrunken-2 and brittle-2 mutants of maize. Biochem Genet. 1976 Aug;14(7-8):547–560. doi: 10.1007/BF00485834. [DOI] [PubMed] [Google Scholar]
  13. Hannah L. C., Tuschall D. M., Mans R. J. Multiple forms of maize endosperm adp-glucose pyrophosphorylase and their control by shrunken-2 and brittle-2. Genetics. 1980 Aug;95(4):961–970. doi: 10.1093/genetics/95.4.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Iglesias A. A., Barry G. F., Meyer C., Bloksberg L., Nakata P. A., Greene T., Laughlin M. J., Okita T. W., Kishore G. M., Preiss J. Expression of the potato tuber ADP-glucose pyrophosphorylase in Escherichia coli. J Biol Chem. 1993 Jan 15;268(2):1081–1086. [PubMed] [Google Scholar]
  15. Kleczkowski L. A., Villand P., Lüthi E., Olsen O. A., Preiss J. Insensitivity of barley endosperm ADP-glucose pyrophosphorylase to 3-phosphoglycerate and orthophosphate regulation. Plant Physiol. 1993 Jan;101(1):179–186. doi: 10.1104/pp.101.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Laughlin M. J., Chantler S. E., Okita T. W. N- and C-terminal peptide sequences are essential for enzyme assembly, allosteric, and/or catalytic properties of ADP-glucose pyrophosphorylase. Plant J. 1998 Apr;14(2):159–168. doi: 10.1046/j.1365-313x.1998.00102.x. [DOI] [PubMed] [Google Scholar]
  18. Laughlin M. J., Payne J. W., Okita T. W. Substrate binding mutants of the higher plant ADP-glucose pyrophosphorylase. Phytochemistry. 1998 Feb;47(4):621–629. doi: 10.1016/s0031-9422(97)00578-5. [DOI] [PubMed] [Google Scholar]
  19. Lin T. P., Caspar T., Somerville C. R., Preiss J. A Starch Deficient Mutant of Arabidopsis thaliana with Low ADPglucose Pyrophosphorylase Activity Lacks One of the Two Subunits of the Enzyme. Plant Physiol. 1988 Dec;88(4):1175–1181. doi: 10.1104/pp.88.4.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lin T. P., Caspar T., Somerville C., Preiss J. Isolation and Characterization of a Starchless Mutant of Arabidopsis thaliana (L.) Heynh Lacking ADPglucose Pyrophosphorylase Activity. Plant Physiol. 1988 Apr;86(4):1131–1135. doi: 10.1104/pp.86.4.1131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Martin C., Smith A. M. Starch biosynthesis. Plant Cell. 1995 Jul;7(7):971–985. doi: 10.1105/tpc.7.7.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nakata P. A., Greene T. W., Anderson J. M., Smith-White B. J., Okita T. W., Preiss J. Comparison of the primary sequences of two potato tuber ADP-glucose pyrophosphorylase subunits. Plant Mol Biol. 1991 Nov;17(5):1089–1093. doi: 10.1007/BF00037149. [DOI] [PubMed] [Google Scholar]
  23. Okita T. W. Is there an alternative pathway for starch synthesis? Plant Physiol. 1992 Oct;100(2):560–564. doi: 10.1104/pp.100.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Plaxton W. C., Preiss J. Purification and Properties of Nonproteolytic Degraded ADPglucose Pyrophosphorylase from Maize Endosperm. Plant Physiol. 1987 Jan;83(1):105–112. doi: 10.1104/pp.83.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Preiss J., Romeo T. Molecular biology and regulatory aspects of glycogen biosynthesis in bacteria. Prog Nucleic Acid Res Mol Biol. 1994;47:299–329. doi: 10.1016/s0079-6603(08)60255-x. [DOI] [PubMed] [Google Scholar]
  26. Smith-White B. J., Preiss J. Comparison of proteins of ADP-glucose pyrophosphorylase from diverse sources. J Mol Evol. 1992 May;34(5):449–464. doi: 10.1007/BF00162999. [DOI] [PubMed] [Google Scholar]
  27. Tsai C. Y., Nelson O. E. Starch-deficient maize mutant lacking adenosine dephosphate glucose pyrophosphorylase activity. Science. 1966 Jan 21;151(3708):341–343. doi: 10.1126/science.151.3708.341. [DOI] [PubMed] [Google Scholar]
  28. Tsai C. Y., Salamini F., Nelson O. E. Enzymes of carbohydrate metabolism in the developing endosperm of maize. Plant Physiol. 1970 Aug;46(2):299–306. doi: 10.1104/pp.46.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wang S. M., Chu B., Lue W. L., Yu T. S., Eimert K., Chen J. adg2-1 represents a missense mutation in the ADPG pyrophosphorylase large subunit gene of Arabidopsis thaliana. Plant J. 1997 May;11(5):1121–1126. doi: 10.1046/j.1365-313x.1997.11051121.x. [DOI] [PubMed] [Google Scholar]

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