Home LiteratureArticle Details
PMID: 8532533 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

mRNA decay in spinach chloroplasts: psbA mRNA degradation is initiated by endonucleolytic cleavages within the coding region.

Nucleic acids research ·Vol. 23 ·No. 23 ·1995-12-11 ·Pages 4885-92

Klaff P

Abstract

The expression of chloroplast genes during leaf development in higher plants is regulated on several levels as transcription, RNA processing and stability, protein stability and turnover. Differential mRNA stability is one major component which contributes to the developmentally controlled accumulation of higher plant chloroplast psbA mRNA, which encodes the D1 protein of photosystem II. To understand the molecular mechanisms of specific mRNA degradation an in vitro mRNA decay system based on lysed chloroplasts from spinach leaves was established. Employing this degradation extract the decay of psbA mRNA was analyzed. Half-life of the psbA mRNA in vitro is dependent on the degradation conditions as the presence of Mg2+, which was found to stabilize the mRNA. Addition of tRNA stabilizes the mRNA and allows the accumulation of distinct degradation intermediates. psbA mRNA derived fragments of the same size were detected in degradation experiments in vitro, in organello and in vivo. 5' ends of the degradation intermediates were identified by primer extension and found to be localized in the 5' part of the coding region. The data indicate a degradation mechanism involving initiation of psbA mRNA decay by specific endonucleolytic cleavage and subsequent exonucleolytic degradation of the fragments. Possible models for cleavage site recognition are discussed.

MeSH Terms
Base Sequence Blotting, Northern Half-Life Molecular Sequence Data RNA, Chloroplast/chemistry,metabolism RNA, Messenger/chemistry,metabolism Spinacia oleracea/genetics,growth & development
Chemicals
RNA, Chloroplast RNA, Messenger
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Klaff P
Heinrich-Heine-Universität Düsseldorf, Institut für Physikalische Biologie, Germany.
References (35)
35 references, click to expand
  1. Control of c-myc mRNA half-life in vitro by a protein capable of binding to a coding region stability determinant.
    Genes Dev. 1992 Apr;6(4):642-54 PMID: 1559612
  2. ard-1: a human gene that reverses the effects of temperature-sensitive and deletion mutations in the Escherichia coli rne gene and encodes an activity producing RNase E-like cleavages.
    Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10591-5 PMID: 7524097
  3. 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. 1990 Oct 25;18(20):6003-10 PMID: 2235482
  4. A 3' stem/loop structure of the Chlamydomonas chloroplast atpB gene regulates mRNA accumulation in vivo.
    Plant Cell. 1991 Mar;3(3):285-97 PMID: 1840911
  5. 5' sequences are important positive and negative determinants of the longevity of Chlamydomonas chloroplast gene transcripts.
    Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1556-60 PMID: 8434017
  6. Synthesis and turnover of photosystem II reaction center protein D1. Ribosome pausing increases during chloroplast development.
    J Biol Chem. 1994 Jul 8;269(27):17918-23 PMID: 8027048
  7. Faithful degradation of soybean rbcS mRNA in vitro.
    Mol Cell Biol. 1994 Apr;14(4):2640-50 PMID: 8139564
  8. Transcriptional and post-transcriptional control of plastid mRNA levels in higher plants.
    Trends Genet. 1988 Sep;4(9):258-63 PMID: 3070872
  9. A cell-free extract from yeast cells for studying mRNA turnover.
    Nucleic Acids Res. 1992 May 25;20(10):2503-10 PMID: 1598208
  10. Influence of Ionic Strength, pH, and Chelation of Divalent Metals on Isolation of Polyribosomes from Tobacco Leaves.
    Plant Physiol. 1976 Jan;57(1):5-10 PMID: 16659423
  11. Plastid run-on transcription. Application to determine the transcriptional regulation of spinach plastid genes.
    J Biol Chem. 1987 Jul 15;262(20):9641-8 PMID: 3597430
  12. Copurification of E. coli RNAase E and PNPase: evidence for a specific association between two enzymes important in RNA processing and degradation.
    Cell. 1994 Mar 11;76(5):889-900 PMID: 7510217
  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. The rate-limiting step in yeast PGK1 mRNA degradation is an endonucleolytic cleavage in the 3'-terminal part of the coding region.
    Mol Cell Biol. 1992 Jul;12(7):2986-96 PMID: 1320194
  15. Specific binding of chloroplast proteins in vitro to the 3' untranslated region of spinach chloroplast petD mRNA.
    Mol Cell Biol. 1991 Sep;11(9):4380-8 PMID: 1908552
  16. Structural analysis of RNA using chemical and enzymatic probing monitored by primer extension.
    Methods Enzymol. 1988;164:481-9 PMID: 2468070
  17. A human RNase E-like activity that cleaves RNA sequences involved in mRNA stability control.
    Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7322-6 PMID: 7638189
  18. Chloroplast gene expression in nuclear, photosynthetic mutants of maize.
    EMBO J. 1986 Jul;5(7):1421-7 PMID: 3743547
  19. Chloroplast gene expression: how plants turn their plastids on.
    Cell. 1989 Jan 27;56(2):161-70 PMID: 2643471
  20. Transcription and RNA stability are important determinants of higher plant chloroplast RNA levels.
    EMBO J. 1987 Jun;6(6):1571-9 PMID: 16453773
  21. H4 histone messenger RNA decay in cell-free extracts initiates at or near the 3' terminus and proceeds 3' to 5'.
    J Mol Biol. 1986 Apr 20;188(4):579-93 PMID: 3525849
  22. Chlorophyll regulates accumulation of the plastid-encoded chlorophyll proteins P700 and D1 by increasing apoprotein stability.
    Plant Physiol. 1994 Mar;104(3):907-16 PMID: 8165261
  23. Nucleotide sequence of the gene for the M(r) 32,000 thylakoid membrane protein from Spinacia oleracea and Nicotiana debneyi predicts a totally conserved primary translation product of M(r) 38,950.
    Proc Natl Acad Sci U S A. 1982 Dec;79(24):7699-703 PMID: 16593262
  24. Nuclease activity associated with mammalian mRNA in its native state: possible basis for selectivity in mRNA decay.
    Mol Cell Biol. 1990 May;10(5):2060-9 PMID: 2325645
  25. Direct evidence for selective modulation of psbA, rpoA, rbcL and 16S RNA stability during barley chloroplast development.
    Plant Mol Biol. 1993 Jun;22(3):447-63 PMID: 8329684
  26. Ribosomes pause at specific sites during synthesis of membrane-bound chloroplast reaction center protein D1.
    J Biol Chem. 1991 Aug 15;266(23):14931-8 PMID: 1869532
  27. Site-specific endonucleolytic cleavages and the regulation of stability of E. coli ompA mRNA.
    Cell. 1988 Mar 25;52(6):893-901 PMID: 3280138
  28. 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
  29. Base-pair probability profiles of RNA secondary structures.
    Comput Appl Biosci. 1992 Aug;8(4):389-99 PMID: 1498695
  30. Changes in Chloroplast mRNA Stability during Leaf Development.
    Plant Cell. 1991 May;3(5):517-529 PMID: 12324602
  31. Dynamic regulation of chloroplast transcription.
    Plant Physiol. 1993 Oct;103(2):309-13 PMID: 8029331
  32. Translation and a 42-nucleotide segment within the coding region of the mRNA encoded by the MAT alpha 1 gene are involved in promoting rapid mRNA decay in yeast.
    Proc Natl Acad Sci U S A. 1990 Apr;87(7):2780-4 PMID: 2181450
  33. Control of plastid gene expression: 3' inverted repeats act as mRNA processing and stabilizing elements, but do not terminate transcription.
    Cell. 1987 Dec 24;51(6):1145-57 PMID: 3690662
  34. Chloroplast gene expression and promoter identification in chloroplast extracts.
    Methods Enzymol. 1986;118:253-70 PMID: 2419734
  35. Identification of endonucleolytic cleavage sites involved in decay of Escherichia coli trxA mRNA.
    J Bacteriol. 1993 Feb;175(4):1043-52 PMID: 7679384
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/23.23.4885
Published
1995-12-11
Pages
4885-92
Language
English
Region
England
NLM ID
0411011
PMCID
PMC307479
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/23.23.4885 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: [email protected]

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