Skip to main content
Log in

A 43 kD light-regulated chloroplast RNA-binding protein interacts with the psbA 5′ non-translated leader RNA

  • Regular Paper
  • Oxygenic Photosynthesis
  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Expression of the chloroplast psbA gene coding for the D1 protein of Photosystem II is subject to regulation at different levels in higher plants, including control of mRNA accumulation and translation. In dicots, the conserved 5′ non-translated leader (5′-UTR) of the psbA mRNA is sufficient to direct the light-dependent translation of the D1 protein. In this report we show that the psbA mRNA 5′-UTR forms a stem-loop structure and binds a 43 kD chloroplast protein (43RNP). Binding of the 43RNP is sensitive to competition with poly(U), but insensitive to high concentrations of tRNA, the RNA homopolymers poly(A), poly(G), poly(C), or poly(A):poly(U) as a double-strand RNA. The 43RNP does not bind efficiently to the psbA mRNA 3′ non-translated region, although the RNA sequence is U-rich and folds into a stem-loop. A deletion mutant of the psbA 5′-UTR RNA in which 5′ sequences of the stem-loop are removed does not affect 43RNP binding. Together, these properties suggest that the 43RNP binds most effectively to a specific single-strand U-rich sequence preceding the AUG start codon in the psbA mRNA. Binding of the 43RNP is not detectable in plastid protein extracts from 5-day-old dark-grown seedlings, but is detectable in light-grown seedlings as well as mature plants in the light and after shifted to the dark. The 43RNP is therefore a candidate for a regulatory RNA-binding protein that may control the accumulation and/or translation of the psbA mRNA during light-dependent seedling development.

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

Abbreviations

DMS:

dimethylsulfate

psb :

Photosystem II genes

RNP:

ribonucleoprotein

UTR:

non-translated leader

UV crosslinking:

ultra-violet light crosslinking

References

  • Adams CC and Stern DB (1990) Control of mRNA stability in chloroplasts by 3′ inverted repeats: Effects of stem and loop mutations on degradation of psbA mRNA in vitro. Nucleic Acids Res 18: 6003–6010

    Google Scholar 

  • Barkan A (1989) Tissue-dependent plastid mRNA splicing in maize: Transcripts from four plastid genes are predominantly unspliced in leaf meristems and roots. Plant Cell 1: 437–445

    Google Scholar 

  • Blum H, Beier H and Gross HJ (1987) Improved silverstaining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8: 93–99

    Google Scholar 

  • Bouvet P and Belasco JG (1992) Control of RNase E-mediated RNA degradation by 5′-terminal basepairing in E. coli. Nature 360: 488–491

    Google Scholar 

  • Chen Q, Adams CC Usack L, Yang J, Monde RA and Stern DB (1995) An AU-rich element in the 3′ untranslated region of the spinach chloroplast petD gene participates in sequence-specific RNA-protein complex formation. Mol Cell Biol. in press

  • Danon A and Mayfield SPY (1991) Light-regulated translational activators: Identification of gene specific mRNA-binding proteins EMBO J 10: 3993–4001

    Google Scholar 

  • Danon A and Mayfield SP (1994a) ADP-dependent phosphorylation regulates RNA-binding in vitro: Implications in light-modulated translation. EMBO J 13: 2227–2235

    Google Scholar 

  • Danon A and Mayfiled SP (1994b) Light-regulated translation of chloroplast messenger RNAs through redox potential. Science 266: 1717–1719

    Google Scholar 

  • Davenloo P, Rosenberg AH, Dunn JJ and Studier FW (1984) Cloning and expression of the gene for bacteriophage T7 RNA polymerase. Proc Natl Acad Sci USA 81: 2035–2039

    Google Scholar 

  • Deng X-W and Gruissem W (1987) Control of plastid gene expression during development: Limited role of transcriptional regulation. Cell 49: 379–387

    Google Scholar 

  • Deng X-W and Gruissem W (1988) Constitutive transcription and regulation of gene expression in non-photosynthetic plastids of higher plants. EMBO J 7: 3301–3308

    Google Scholar 

  • Ehresmann C, Baudin F, Mougel M, Romby P, Ebel J-P and Ehresmann B (1987) Probing the structure of RNA in solution. Nucl Acids Res 15: 9109–9128

    Google Scholar 

  • Eichacker LA, Söll J, Lauterback P, Rüdiger P, Klein RR and Mullet JR (1990) In vitro synthesis of chlorophyll A in the dark triggers accumulation of chlorophyll A apoproteins in barley etioplasts. J Biol Chem 265: 13566–13571

    Google Scholar 

  • Franzetti B, Carol P and Mache R (1992) Characterization and RNA-binding properties of a chloroplast S1-like ribosomal protein. J Biol Chem 267: 19075–19081

    Google Scholar 

  • Fromm H, Devic M, Fluhr R and Edelman M (1985) Control of psbA gene expression: In mature Spirodela chloroplasts light regulation of 32-kD protein synthesis is independent of transcript level. EMBO J 4: 291–295

    Google Scholar 

  • Greenberg JR (1980) Proteins crosslinked to messenger RNA by irradiating polyribosomes with ultraviolet light. Nucleic Acids Res 8: 5685–5701

    Google Scholar 

  • Gruissem W, Greenberg BM, Zurawski G and Hallick RB (1986) Chloroplast gene expression and promotor identification in chloroplasts extracts. Meth Enzymol 118: 253–270

    Google Scholar 

  • Gruissem W, Deng X-W, Jones H, Stern D, Tonkyn J and Zurawski G (1987) Transcriptional and post-transcriptional regulation of chloroplast gene expression. In: vonWettstein D and Chua N-H (eds) Plant Molecular Biology, pp 135–148. Plenum Press, New York

    Google Scholar 

  • Gruissem W, Barkan A, Deng X-W and Stern D (1988) Transcriptional and post-transcriptional control of plastid mRNA levels in higher plants. Trends Gen 4: 258–263

    Google Scholar 

  • Gruissem W (1989) Chloroplast gene expression: how plants turn their plastids on. Cell 56: 161–170

    Google Scholar 

  • Gruissem W and Schuster G (1993) Control of mRNA stability and degradation in organelles. In: Brawerman G and Belasco J (eds) Control of mRNA stability, pp 329–365. Academic Press, New York

    Google Scholar 

  • Gruissem W and Tonkyn JC (1993) Control mechanisms in plastid gene expression. Crit Rev Plant Sci 12: 19–55

    Google Scholar 

  • Herrmann RG, Westhoff P, Alt J, Tittgen J and Nelson N (1985) Thylakoid membrane proteins and their genes. In: vanVloten-Doting L, Groot GSP and Hall TC (eds) Molecular Form and Function of the Plant Genome, pp 233–256. Plenum Press, New York

    Google Scholar 

  • Jackson AO and Larkins BA (1976) Influence of ionic strength, pH, and chelation of divalent metals on the isolation of polyribosomes from tobacco leaves. Plant Physiol 57: 5–10

    Google Scholar 

  • Kim J, Klein PG and Mullet JE (1991) Ribosomes pause at specific sites during synthesis of membrane-bound chloroplast reaction center protein D1. J Biol Chem 266: 14931–14938

    Google Scholar 

  • Kim J, Klein PG and Mullet JE (1994) Synthesis and turnover of Photosystem II reaction center protein D1. J Biol Chem 269: 17918–17923

    Google Scholar 

  • Klaff P and Gruissem W (1991) Changes in chloroplast mRNA stability during leaf development. Plant Cell 3: 517–529

    Google Scholar 

  • Klein RR and Mullet JE (1986) Regulation of chloroplast-encoded chlorophyll-binding protein translation during higher plant chloroplast biogenesis. J Biol Chem 261: 11138–11145

    Google Scholar 

  • Klein RR and Mullet JE (1990) Light-induced transcription of chloroplast genes. psbA transcription is differentially enhanced in illuminated barley. J Biol Chem 265: 1895–1902

    Google Scholar 

  • Klein RR, Mason HS and Mullet JE (1988) 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 106: 289–301

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  • Li Y and Sugiura M (1990) Three distinct ribonucleoproteins from tobacco chloroplasts: Each contains a unique amino terminal acidic domain and two ribonucleoprotein consensus motives. EMBO J 9: 3059–3066

    Google Scholar 

  • Lundberg U, vonGabain A and Melefors OE (1990) Cleavages in the 5′ region of the ompA and bla mRNA control stability: Studies with an E. coli mutant altering mRNA stability and a novel endoribonuclease. EMBO J 9: 2731–2741

    Google Scholar 

  • Mayford M and Weisblum B (1989) Conformational alterations in the ermC transcript in vivo during induction. EMBO J 8: 4307–4314

    Google Scholar 

  • Mullet JE (1988) Chloroplast development and gene expression. Annu Rev Plant Physiol Plant Mol Biol 39: 475–502

    Google Scholar 

  • Mullet JE and Klein RR (1987) Transcription and RNA stability are important determinants of higher plant chloroplast RNA levels. EMBO J 6: 1571–1579

    Google Scholar 

  • Mullet JE, Klein PG and Klein RR (1990) Chlorophyll regulates accumulation of the plastid-encoded apoproteins CP43 and D1 by increasing apoprotein stability. Proc Natl Acad Sci USA 87: 4038–4042

    Google Scholar 

  • Nickelsen J and Link G (1989) Interaction of a 3′ RNA region of the mustard trnK gene with chloroplast proteins. Nucleic Acids Res 23: 9637–9648

    Google Scholar 

  • Nickelsen J and Link G (1991) RNA-protein interactions at transcript 3′ ends and evidence for trnK-psbA cotranscription in mustard chloroplasts. Mol Gen Genet 228: 89–96

    Google Scholar 

  • Nickelsen J, vanDillewijn J, Rahire M and Rochaix J-D (1994) Determinants for stability of the chloroplast psbD RNA are located within the short leader region in Chlamydomonas reinhardii. EMBO J 13: 3182–3191

    Google Scholar 

  • Piechulla B, Pichersky E, Cashmore AR and Gruissem W (1986) Expression of nuclear and plastid genes for photosynthesis-specific proteins during tomato fruit development and ripening. Plant Mol Biol 7: 367–376

    Google Scholar 

  • Rapp JC, Baumgartner BJ and Mullet JE (1992) Quantitative analysis of transcription and RNA levels of 15 barley chloroplast genes—transcription rates and messenger RNA levels vary over 300-fold —predicted messenger RNA stabilities vary 30-fold. J Biol Chem 267: 21404–21411

    Google Scholar 

  • Riesselmann S and Piechulla B (1992) Diurnal and circadian light-harvesting complex and quinone B-binding protein synthesis in leaves of tomato (Lycopersicon esculentum). Plant Physiol 100: 1840–1845

    Google Scholar 

  • Rochaix J-D (1992) Post-transcriptional steps in the expression of chloroplast genes. Annu Rev Cell Biol 8: 1–28

    Google Scholar 

  • Sakamoto W, Kindle KL and Stern DB (1993) in vivo analysis of Chlamydomonas chloroplast petD expression using stable trans-formation of beta-glucuronidase translational fusions. Proc Natl Acad Sci USA 90: 497–501

    Google Scholar 

  • Salvador ML, Klein U and Bogorad L (1993) 5′ sequences are important positive and negative determinants of the longevity of Chlamydomonas chloroplast gene transcripts. Proc Natl Acad Sci USA 90: 1556–1560

    Google Scholar 

  • Schmitz M and Steger G (1992) Base-pair probability profiles of RNA secondary structure. Comp Appl Biol Sci 8: 389–399

    Google Scholar 

  • Schuster G and Gruissem W (1991) Chloroplast 3′ end processing requires a nuclear encoded RNA-binding protein. EMBO J 10: 1493–1503

    Google Scholar 

  • Staub JM and Maliga P (1993) Accumulation of D1 polypeptide in tobacco plastids via the untranslated region of the psbA mRNA. EMBO J 12: 601–606

    Google Scholar 

  • Staub JM and Maliga P (1994) Translation of psbA mRNA is regulated by light via the 5′ untranslated region in tobacco plastids. Plant J 6: 547–553

    Google Scholar 

  • Stebbins-Boaz B and Gerbi SA (1991) Structural analysis of the peptidyl transferase region in ribosomal RNA of the eukaryote Xenopus laevis. J Mol Biol 217: 93–112

    Google Scholar 

  • Stern DB and Gruissem W (1987) Control of plastid gene expression: 3′ inverted repeats act as mRNA processing signals and stabilizing elements, but do not terminate transcription. Cell 51: 1145–1157

    Google Scholar 

  • Stern DB, Jones H and Gruissem W (1989) Function of plastid mRNA 3′ inverted repeats: RNA stabilization and gene specific protein binding. J Biol Chem 264: 18742–18750

    Google Scholar 

  • Stern DB, Radwanski ER and Kindle K (1991) A 3′ stem-loop structure of the Chlamydomonas chloroplast atpB gene regulates mRNA accumulation in vivo. Plant Cell 3: 285–297

    Google Scholar 

  • Tang CK and Draper DE (1989) Unusual MRNA pseudoknot structure is recognized by a protein translational repressor. Cell 57: 531–536

    Google Scholar 

  • Theil EC (1990) Regulation of ferritin and transferrin receptor mRNAs. J Biol Chem 265: 4771–4774

    Google Scholar 

  • Tiller K and Link G (1993) Phosphorylation and dephosphorylation affect functional characteristics of chloroplast and etioplast transcription systems from mustard (Sinapis alba L.). EMBO J 12: 1745–1753

    Google Scholar 

  • Tonkyn JC, Deng X-W and Gruissem W (1992) Regulation of plastid gene expression during photosynthetic stress. Plant Physiol 99: 1406–1415

    Google Scholar 

  • Walter P and Blobel G (1982) Disassembly and reconstitution of signal recognition particle. Cell 34: 525–533

    Google Scholar 

  • Zerges W and Rochaix J-D (1994) The 5′ leader of a chloroplast mRNA mediates the translational requirements for two nucleusencoded functions in Chlamydomonas reinhardii. Mol Cell Biol 14: 5268–5277

    Google Scholar 

  • Zurawski G, Bohnert HJ, Whitfeld PR and Bottomley W (1982) Nucleotide sequence of the gene for the Mr 32,000 thylacoid membrane protein from Spinacea oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of Mr 38,950. Proc Natl Acad Sci USA 79: 7699–7703

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Klaff, P., Gruissem, W. A 43 kD light-regulated chloroplast RNA-binding protein interacts with the psbA 5′ non-translated leader RNA. Photosynth Res 46, 235–248 (1995). https://doi.org/10.1007/BF00020436

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00020436

Key words

Navigation