Skip to main content
The Plant Cell logoLink to The Plant Cell
. 1999 May;11(5):849–864. doi: 10.1105/tpc.11.5.849

BUNDLE SHEATH DEFECTIVE2, a novel protein required for post-translational regulation of the rbcL gene of maize.

T P Brutnell 1, R J Sawers 1, A Mant 1, J A Langdale 1
PMCID: PMC144220  PMID: 10330470

Abstract

The Bundle sheath defective2 (Bsd2) gene is required for ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) accumulation in maize. Using a Mutator transposable element as a molecular probe, we identified a tightly linked restriction fragment length polymorphism that cosegregated with the bsd2-conferred phenotype. This fragment was cloned, and sequences flanking the Mutator insertion were used to screen a maize leaf cDNA library. Using a full-length cDNA clone isolated in this screen, we show that an abundant 0.6-kb transcript could be detected in wild-type plants but not in bsd2-m1 plants. This 0.6-kb transcript accumulated to low levels in plants carrying an allele derived from bsd2-m1 that conditions a less severe mutant phenotype. Taken together, these data strongly suggest that we have cloned the Bsd2 gene. Sequence analysis of the full-length cDNA clone revealed a chloroplast targeting sequence and a region of homology shared between BSD2 and the DnaJ class of molecular chaperones. This region of homology is limited to a cysteine-rich Zn binding domain in DnaJ believed to play a role in protein-protein interactions. We show that BSD2 is targeted to the chloroplast but is not involved in general photosynthetic complex assembly or protein import. In bsd2 mutants, we could not detect the Rubisco protein, but the chloroplast-encoded Rubisco large subunit transcript (rbcL) was abundant and associated with polysomes in both bundle sheath and mesophyll cells. By characterizing Bsd2 expression patterns and analyzing the bsd2-conferred phenotype, we propose a model for BSD2 in the post-translational regulation of rbcL in maize.

Full Text

The Full Text of this article is available as a PDF (767.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  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. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997 Sep 1;25(17):3389–3402. doi: 10.1093/nar/25.17.3389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Banecki B., Liberek K., Wall D., Wawrzynów A., Georgopoulos C., Bertoli E., Tanfani F., Zylicz M. Structure-function analysis of the zinc finger region of the DnaJ molecular chaperone. J Biol Chem. 1996 Jun 21;271(25):14840–14848. doi: 10.1074/jbc.271.25.14840. [DOI] [PubMed] [Google Scholar]
  4. Banecki B., Zylicz M. Real time kinetics of the DnaK/DnaJ/GrpE molecular chaperone machine action. J Biol Chem. 1996 Mar 15;271(11):6137–6143. doi: 10.1074/jbc.271.11.6137. [DOI] [PubMed] [Google Scholar]
  5. Barkan A., Martienssen R. A. Inactivation of maize transposon Mu suppresses a mutant phenotype by activating an outward-reading promoter near the end of Mu1. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3502–3506. doi: 10.1073/pnas.88.8.3502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barkan A. Nuclear Mutants of Maize with Defects in Chloroplast Polysome Assembly Have Altered Chloroplast RNA Metabolism. Plant Cell. 1993 Apr;5(4):389–402. doi: 10.1105/tpc.5.4.389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brutnell T. P., Dellaporta S. L. Somatic inactivation and reactivation of Ac associated with changes in cytosine methylation and transposase expression. Genetics. 1994 Sep;138(1):213–225. doi: 10.1093/genetics/138.1.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chandler V. L., Hardeman K. J. The Mu elements of Zea mays. Adv Genet. 1992;30:77–122. doi: 10.1016/s0065-2660(08)60319-3. [DOI] [PubMed] [Google Scholar]
  9. Checa S. K., Viale A. M. The 70-kDa heat-shock protein/DnaK chaperone system is required for the productive folding of ribulose-biphosphate carboxylase subunits in Escherichia coli. Eur J Biochem. 1997 Sep 15;248(3):848–855. doi: 10.1111/j.1432-1033.1997.00848.x. [DOI] [PubMed] [Google Scholar]
  10. Chomet P., Lisch D., Hardeman K. J., Chandler V. L., Freeling M. Identification of a regulatory transposon that controls the Mutator transposable element system in maize. Genetics. 1991 Sep;129(1):261–270. doi: 10.1093/genetics/129.1.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Deng X. W., Gruissem W. Control of plastid gene expression during development: the limited role of transcriptional regulation. Cell. 1987 May 8;49(3):379–387. doi: 10.1016/0092-8674(87)90290-x. [DOI] [PubMed] [Google Scholar]
  12. Fowler J. E., Muehlbauer G. J., Freeling M. Mosaic analysis of the liguleless3 mutant phenotype in maize by coordinate suppression of mutator-insertion alleles. Genetics. 1996 May;143(1):489–503. doi: 10.1093/genetics/143.1.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gillham N. W., Boynton J. E., Hauser C. R. Translational regulation of gene expression in chloroplasts and mitochondria. Annu Rev Genet. 1994;28:71–93. doi: 10.1146/annurev.ge.28.120194.000443. [DOI] [PubMed] [Google Scholar]
  14. Greene B., Walko R., Hake S. Mutator insertions in an intron of the maize knotted1 gene result in dominant suppressible mutations. Genetics. 1994 Dec;138(4):1275–1285. doi: 10.1093/genetics/138.4.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gutteridge S., Gatenby A. A. Rubisco Synthesis, Assembly, Mechanism, and Regulation. Plant Cell. 1995 Jul;7(7):809–819. doi: 10.1105/tpc.7.7.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hall L. N., Rossini L., Cribb L., Langdale J. A. GOLDEN 2: a novel transcriptional regulator of cellular differentiation in the maize leaf. Plant Cell. 1998 Jun;10(6):925–936. doi: 10.1105/tpc.10.6.925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hartl F. U. Molecular chaperones in cellular protein folding. Nature. 1996 Jun 13;381(6583):571–579. doi: 10.1038/381571a0. [DOI] [PubMed] [Google Scholar]
  18. Kelley W. L. The J-domain family and the recruitment of chaperone power. Trends Biochem Sci. 1998 Jun;23(6):222–227. doi: 10.1016/s0968-0004(98)01215-8. [DOI] [PubMed] [Google Scholar]
  19. Kim J., Klein P. G., Mullet J. E. Synthesis and turnover of photosystem II reaction center protein D1. Ribosome pausing increases during chloroplast development. J Biol Chem. 1994 Jul 8;269(27):17918–17923. [PubMed] [Google Scholar]
  20. Klaff P., Gruissem W. Changes in Chloroplast mRNA Stability during Leaf Development. Plant Cell. 1991 May;3(5):517–529. doi: 10.1105/tpc.3.5.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Klein R. R., Mason H. S., Mullet J. E. Light-regulated translation of chloroplast proteins. I. Transcripts of psaA-psaB, psbA, and rbcL are associated with polysomes in dark-grown and illuminated barley seedlings. J Cell Biol. 1988 Feb;106(2):289–301. doi: 10.1083/jcb.106.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kubicki A., Steinmüller K., Westhoff P. Differential transcription of plastome-encoded genes in the mesophyll and bundle-sheath chloroplasts of the monocotyledonous NADP-malic enzyme-type C4 plants maize and Sorghum. Plant Mol Biol. 1994 Jul;25(4):669–679. doi: 10.1007/BF00029605. [DOI] [PubMed] [Google Scholar]
  23. Langdale J. A., Metzler M. C., Nelson T. The argentia mutation delays normal development of photosynthetic cell-types in Zea mays. Dev Biol. 1987 Jul;122(1):243–255. doi: 10.1016/0012-1606(87)90349-6. [DOI] [PubMed] [Google Scholar]
  24. Langdale J. A., Rothermel B. A., Nelson T. Cellular pattern of photosynthetic gene expression in developing maize leaves. Genes Dev. 1988 Jan;2(1):106–115. doi: 10.1101/gad.2.1.106. [DOI] [PubMed] [Google Scholar]
  25. Langdale J. A., Zelitch I., Miller E., Nelson T. Cell position and light influence C4 versus C3 patterns of photosynthetic gene expression in maize. EMBO J. 1988 Dec 1;7(12):3643–3651. doi: 10.1002/j.1460-2075.1988.tb03245.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Langer T., Lu C., Echols H., Flanagan J., Hayer M. K., Hartl F. U. Successive action of DnaK, DnaJ and GroEL along the pathway of chaperone-mediated protein folding. Nature. 1992 Apr 23;356(6371):683–689. doi: 10.1038/356683a0. [DOI] [PubMed] [Google Scholar]
  27. Martienssen R., Barkan A., Taylor W. C., Freeling M. Somatically heritable switches in the DNA modification of Mu transposable elements monitored with a suppressible mutant in maize. Genes Dev. 1990 Mar;4(3):331–343. doi: 10.1101/gad.4.3.331. [DOI] [PubMed] [Google Scholar]
  28. Martineau B., Taylor W. C. Photosynthetic gene expression and cellular differentiation in developing maize leaves. Plant Physiol. 1985 Jun;78(2):399–404. doi: 10.1104/pp.78.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mould R. M., Shackleton J. B., Robinson C. Transport of proteins into chloroplasts. Requirements for the efficient import of two lumenal oxygen-evolving complex proteins into isolated thylakoids. J Biol Chem. 1991 Sep 15;266(26):17286–17289. [PubMed] [Google Scholar]
  30. Nakai K., Kanehisa M. A knowledge base for predicting protein localization sites in eukaryotic cells. Genomics. 1992 Dec;14(4):897–911. doi: 10.1016/S0888-7543(05)80111-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nelson T., Harpster M. H., Mayfield S. P., Taylor W. C. Light-regulated gene expression during maize leaf development. J Cell Biol. 1984 Feb;98(2):558–564. doi: 10.1083/jcb.98.2.558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Reinbothe S., Reinbothe C., Lebedev N., Apel K. PORA and PORB, Two Light-Dependent Protochlorophyllide-Reducing Enzymes of Angiosperm Chlorophyll Biosynthesis. Plant Cell. 1996 May;8(5):763–769. doi: 10.1105/tpc.8.5.763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rochaix J. D. Post-transcriptional steps in the expression of chloroplast genes. Annu Rev Cell Biol. 1992;8:1–28. doi: 10.1146/annurev.cb.08.110192.000245. [DOI] [PubMed] [Google Scholar]
  34. Rodermel S. R., Abbott M. S., Bogorad L. Nuclear-organelle interactions: nuclear antisense gene inhibits ribulose bisphosphate carboxylase enzyme levels in transformed tobacco plants. Cell. 1988 Nov 18;55(4):673–681. doi: 10.1016/0092-8674(88)90226-7. [DOI] [PubMed] [Google Scholar]
  35. Rodermel S., Haley J., Jiang C. Z., Tsai C. H., Bogorad L. A mechanism for intergenomic integration: abundance of ribulose bisphosphate carboxylase small-subunit protein influences the translation of the large-subunit mRNA. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):3881–3885. doi: 10.1073/pnas.93.9.3881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Roth R., Hall L. N., Brutnell T. P., Langdale J. A. bundle sheath defective2, a Mutation That Disrupts the Coordinated Development of Bundle Sheath and Mesophyll Cells in the Maize Leaf. Plant Cell. 1996 May;8(5):915–927. doi: 10.1105/tpc.8.5.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Santel H. J., Apel K. The protochlorophyllide holochrome of barley (Hordeum vulgare L.). The effect of light on the NADPH:protochlorophyllide oxidoreductase. Eur J Biochem. 1981 Nov;120(1):95–103. doi: 10.1111/j.1432-1033.1981.tb05674.x. [DOI] [PubMed] [Google Scholar]
  38. Schlicher T., Soll J. Chloroplastic isoforms of DnaJ and GrpE in pea. Plant Mol Biol. 1997 Jan;33(1):181–185. doi: 10.1023/a:1005784115363. [DOI] [PubMed] [Google Scholar]
  39. Schmidt G. W., Mishkind M. L. Rapid degradation of unassembled ribulose 1,5-bisphosphate carboxylase small subunits in chloroplasts. Proc Natl Acad Sci U S A. 1983 May;80(9):2632–2636. doi: 10.1073/pnas.80.9.2632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Schulz R., Steinmüller K., Klaas M., Forreiter C., Rasmussen S., Hiller C., Apel K. Nucleotide sequence of a cDNA coding for the NADPH-protochlorophyllide oxidoreductase (PCR) of barley (Hordeum vulgare L.) and its expression in Escherichia coli. Mol Gen Genet. 1989 Jun;217(2-3):355–361. doi: 10.1007/BF02464904. [DOI] [PubMed] [Google Scholar]
  41. Schäffner A. R., Sheen J. Maize rbcS promoter activity depends on sequence elements not found in dicot rbcS promoters. Plant Cell. 1991 Sep;3(9):997–1012. doi: 10.1105/tpc.3.9.997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sheen J. Y., Bogorad L. Differential expression of six light-harvesting chlorophyll a/b binding protein genes in maize leaf cell types. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7811–7815. doi: 10.1073/pnas.83.20.7811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sheen J. Y., Bogorad L. Differential expression of the ribulose bisphosphate carboxylase large subunit gene in bundle sheath and mesophyll cells of developing maize leaves is influenced by light. Plant Physiol. 1985 Dec;79(4):1072–1076. doi: 10.1104/pp.79.4.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sheen J. Y., Bogorad L. Expression of the ribulose-1,5-bisphosphate carboxylase large subunit gene and three small subunit genes in two cell types of maize leaves. EMBO J. 1986 Dec 20;5(13):3417–3422. doi: 10.1002/j.1460-2075.1986.tb04663.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Smith LH, Langdale JA, Chollet R. A functional calvin cycle is not indispensable for the light activation of C4 phosphoenolpyruvate carboxylase kinase and its target enzyme in the maize mutant bundle sheath defective2-mutable1 . Plant Physiol. 1998 Sep;118(1):191–197. doi: 10.1104/pp.118.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Spreitzer R. J., Goldschmidt-Clermont M., Rahire M., Rochaix J. D. Nonsense mutations in the Chlamydomonas chloroplast gene that codes for the large subunit of ribulosebisphosphate carboxylase/oxygenase. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5460–5464. doi: 10.1073/pnas.82.16.5460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Szabo A., Korszun R., Hartl F. U., Flanagan J. A zinc finger-like domain of the molecular chaperone DnaJ is involved in binding to denatured protein substrates. EMBO J. 1996 Jan 15;15(2):408–417. [PMC free article] [PubMed] [Google Scholar]
  48. Szabo A., Langer T., Schröder H., Flanagan J., Bukau B., Hartl F. U. The ATP hydrolysis-dependent reaction cycle of the Escherichia coli Hsp70 system DnaK, DnaJ, and GrpE. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10345–10349. doi: 10.1073/pnas.91.22.10345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Troutt A. B., McHeyzer-Williams M. G., Pulendran B., Nossal G. J. Ligation-anchored PCR: a simple amplification technique with single-sided specificity. Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9823–9825. doi: 10.1073/pnas.89.20.9823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Viret J. F., Mabrouk Y., Bogorad L. Transcriptional photoregulation of cell-type-preferred expression of maize rbcS-m3: 3' and 5' sequences are involved. Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8577–8581. doi: 10.1073/pnas.91.18.8577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wang J. L., Turgeon R., Carr J. P., Berry J. O. Carbon Sink-to-Source Transition Is Coordinated with Establishment of Cell-Specific Gene Expression in a C4 Plant. Plant Cell. 1993 Mar;5(3):289–296. doi: 10.1105/tpc.5.3.289. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES