Home LiteratureArticle Details
PMID: 8612593 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cis-acting elements and trans-acting factors for accurate translation of chloroplast psbA mRNAs: development of an in vitro translation system from tobacco chloroplasts.

The EMBO journal ·Vol. 15 ·No. 7 ·1996-04-01 ·Pages 1687-95

Hirose T, Sugiura M

Abstract

Translational regulation is an important step of gene expression in chloroplasts. To analyze biochemical mechanisms of translational regulation unique to higher plant chloroplasts, an in vitro translation system has been developed from tobacco chloroplasts. Conditions for chloroplast extraction and the in vitro translation reaction have been optimized with a tobacco psbA-lacZ fusion mRNA. The in vitro system supports accurate translation of a variety of chloroplasts mRNAs. Using a series of mutant psbA mRNAs, we showed that three elements within the 5'-untranslated region of the mRNA are required for translation. Two of them are complementary to the 3'-terminus of chloroplast 16S rRNA (termed RBS1 and RBS2) and the other is an AU-rich sequence (UAAAUAAA) located between RBS1 and RBS2 and is termed the AU box. mRNA competition experiments using the in vitro translation reaction and gel mobility shift assays revealed the existence of a trans-acting factor(s) for translation and its possible interaction with the AU box. We propose a model for the initiation of psbA translation whereby RBS1 and RBS2 bind cooperatively to the 3'-end of 16S rRNA resulting in looping out of the AU box, which facilitates the interaction of a trans-acting factor(s).

MeSH Terms
Base Sequence Binding Sites Chloroplasts/genetics Cloning, Molecular Gene Expression Regulation, Plant Lac Operon Models, Genetic Molecular Sequence Data Mutation Photosynthetic Reaction Center Complex Proteins/genetics Photosystem II Protein Complex Plants, Toxic Protein Biosynthesis RNA, Messenger/genetics,metabolism RNA, Plant/genetics,metabolism RNA, Ribosomal, 16S/genetics Ribosomes/metabolism Tobacco/genetics
Chemicals
Photosynthetic Reaction Center Complex Proteins Photosystem II Protein Complex RNA, Messenger RNA, Plant RNA, Ribosomal, 16S
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Hirose T
Center for Gene Research, Nagoya University, Japan.
Sugiura M
References (44)
44 references, click to expand
  1. Light regulated translational activators: identification of chloroplast gene specific mRNA binding proteins.
    EMBO J. 1991 Dec;10(13):3993-4001 PMID: 1721866
  2. Proteins encoded by a complex chloroplast transcription unit are each translated from both monocistronic and polycistronic mRNAs.
    EMBO J. 1988 Sep;7(9):2637-44 PMID: 2460341
  3. Large subunit ribulosebisphosphate carboxylase messenger RNA from Euglena chloroplasts.
    Proc Natl Acad Sci U S A. 1976 Mar;73(3):722-6 PMID: 815906
  4. Accumulation of D1 polypeptide in tobacco plastids is regulated via the untranslated region of the psbA mRNA.
    EMBO J. 1993 Feb;12(2):601-6 PMID: 8440249
  5. Ribosome-binding sites on chloroplast rbcL and psbA mRNAs and light-induced initiation of D1 translation.
    Plant Mol Biol. 1994 Jun;25(3):437-48 PMID: 8049369
  6. The chloroplast genome.
    Plant Mol Biol. 1992 May;19(1):149-68 PMID: 1600166
  7. Spinach chloroplast mRNA for a 32 000 dalton polypeptide: size and localization on the physical map of the chloroplast DNA.
    Biochim Biophys Acta. 1980 Dec 11;610(2):297-310 PMID: 7213628
  8. Stable transformation of plastids in higher plants.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8526-30 PMID: 11607112
  9. ADP-dependent phosphorylation regulates RNA-binding in vitro: implications in light-modulated translation.
    EMBO J. 1994 May 1;13(9):2227-35 PMID: 8187775
  10. An additional ribosome-binding site on mRNA of highly expressed genes and a bifunctional site on the colicin fragment of 16S rRNA from Escherichia coli: important determinants of the efficiency of translation-initiation.
    Nucleic Acids Res. 1989 Apr 25;17(8):2973-85 PMID: 2657653
  11. Characterization and RNA-binding properties of a chloroplast S1-like ribosomal protein.
    J Biol Chem. 1992 Sep 25;267(27):19075-81 PMID: 1527032
  12. Effect of the secondary structure in the Euglena gracilis chloroplast ribulose-bisphosphate carboxylase/oxygenase messenger RNA on translational initiation.
    J Biol Chem. 1994 Mar 11;269(10):7501-8 PMID: 8125969
  13. In vivo analysis of Chlamydomonas chloroplast petD gene expression using stable transformation of beta-glucuronidase translational fusions.
    Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):497-501 PMID: 8421681
  14. Translation of the psbA mRNA of Chlamydomonas reinhardtii requires a structured RNA element contained within the 5' untranslated region.
    J Cell Biol. 1994 Dec;127(6 Pt 1):1537-45 PMID: 7798310
  15. Translational regulation of gene expression in chloroplasts and mitochondria.
    Annu Rev Genet. 1994;28:71-93 PMID: 7893142
  16. High-frequency plastid transformation in tobacco by selection for a chimeric aadA gene.
    Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):913-7 PMID: 8381537
  17. Translation of psbA mRNA is regulated by light via the 5'-untranslated region in tobacco plastids.
    Plant J. 1994 Oct;6(4):547-53 PMID: 7987413
  18. Initiation complex formation on Euglena chloroplast 30S subunits in the presence of natural mRNAs.
    Nucleic Acids Res. 1989 Dec 11;17(23):9735-47 PMID: 2690007
  19. Light-regulated translation of chloroplast messenger RNAs through redox potential.
    Science. 1994 Dec 9;266(5191):1717-9 PMID: 7992056
  20. The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression.
    EMBO J. 1986 Sep;5(9):2043-2049 PMID: 16453699
  21. Complete RNA editing of unspliced and dicistronic transcripts of the intron-containing reading frame IRF170 from maize chloroplasts.
    Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2295-9 PMID: 7545915
  22. Prokaryotic translation: the interactive pathway leading to initiation.
    Trends Genet. 1994 Nov;10(11):402-7 PMID: 7809946
  23. Post-transcriptional steps in the expression of chloroplast genes.
    Annu Rev Cell Biol. 1992;8:1-28 PMID: 1282351
  24. A methyl jasmonate-induced shift in the length of the 5' untranslated region impairs translation of the plastid rbcL transcript in barley.
    EMBO J. 1993 Apr;12(4):1505-12 PMID: 8467800
  25. In vitro expression of chloroplast genes in lysates of higher plant chloroplasts.
    Proc Natl Acad Sci U S A. 1985 Jun;82(12):3983-7 PMID: 3858855
  26. The complete nucleotide sequence of 16S ribosomal RNA gene from tobacco chloroplasts.
    Gene. 1982 Feb;17(2):213-8 PMID: 7044901
  27. Chloroplast ribosomes and protein synthesis.
    Microbiol Rev. 1994 Dec;58(4):700-54 PMID: 7854253
  28. Chloroplast transformation in Chlamydomonas with high velocity microprojectiles.
    Science. 1988 Jun 10;240(4858):1534-8 PMID: 2897716
  29. Determinants for stability of the chloroplast psbD RNA are located within its short leader region in Chlamydomonas reinhardtii.
    EMBO J. 1994 Jul 1;13(13):3182-91 PMID: 8039511
  30. Molecular genetics of chloroplast ribosomal proteins.
    Trends Biochem Sci. 1993 May;18(5):177-81 PMID: 8328017
  31. Ribosome-messenger recognition: mRNA target sites for ribosomal protein S1.
    Nucleic Acids Res. 1991 Jan 11;19(1):155-62 PMID: 2011495
  32. The 5' leader of a chloroplast mRNA mediates the translational requirements for two nucleus-encoded functions in Chlamydomonas reinhardtii.
    Mol Cell Biol. 1994 Aug;14(8):5268-77 PMID: 8035805
  33. Translation of chloroplast-encoded mRNA: potential initiation and termination signals.
    Nucleic Acids Res. 1989 Mar 11;17(5):2057-80 PMID: 2928114
  34. ATP-dependent protein synthesis in isolated pea chloroplasts. Evidence for accumulation of a translation intermediate of the D1 protein.
    FEBS Lett. 1993 Feb 8;317(1-2):57-61 PMID: 8428635
  35. Function of the Chlamydomonas reinhardtii petd 5' untranslated region in regulating the accumulation of subunit IV of the cytochrome b6/f complex.
    Plant J. 1994 Oct;6(4):503-12 PMID: 7987409
  36. A uridine-rich sequence required for translation of prokaryotic mRNA.
    Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2605-9 PMID: 1372983
  37. Analysis of the translational initiation region on the Euglena gracilis chloroplast ribulose-bisphosphate carboxylase/oxygenase (rbcL) messenger RNA.
    J Biol Chem. 1994 Mar 11;269(10):7494-500 PMID: 8125968
  38. A nuclear mutation in maize blocks the processing and translation of several chloroplast mRNAs and provides evidence for the differential translation of alternative mRNA forms.
    EMBO J. 1994 Jul 1;13(13):3170-81 PMID: 8039510
  39. RNA editing of tobacco petB mRNAs occurs both in chloroplasts and non-photosynthetic proplastids.
    Plant Mol Biol. 1994 Oct;26(1):509-13 PMID: 7948899
  40. Maize plastid gene expressed during photoregulated development.
    Proc Natl Acad Sci U S A. 1978 Jul;75(7):3060-4 PMID: 16592541
  41. Constitutive transcription and regulation of gene expression in non-photosynthetic plastids of higher plants.
    EMBO J. 1988 Nov;7(11):3301-8 PMID: 16453858
  42. Protein synthesis in chloroplasts. V. Translation of messenger RNA for the large subunit of fraction I protein in a heterologous cell-free system.
    J Mol Biol. 1975 Jan 5;91(1):67-77 PMID: 1102697
  43. Initiation codon mutations in the Chlamydomonas chloroplast petD gene result in temperature-sensitive photosynthetic growth.
    EMBO J. 1993 Sep;12(9):3627-35 PMID: 8253086
  44. RNA editing in maize chloroplasts is a processing step independent of splicing and cleavage to monocistronic mRNAs.
    Plant J. 1993 Oct;4(4):621-9 PMID: 8252066
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1996-04-01
Pages
1687-95
Language
English
Region
England
NLM ID
8208664
PMCID
PMC450080
Subset
IM
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2