Overexpression of the clpP 5′-untranslated region in a chimeric context causes a mutant phenotype, suggesting competition for a clpP-specific RNA maturation factor in tobacco chloroplasts

Hiroshi Kuroda, Pal Maliga

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The plastid ribosomal RNA (rrn) operon promoter was fused with DNA segments encoding the leader sequence (5′untranslated region [UTR]) of plastid mRNAs to compare their efficiency in mediating translation of a bacterial protein neomycin phosphotransferase (NPTII) in tobacco (Nicotiana tabacum) chloroplasts. In young leaves, NPTII accumulated at 0.26% and 0.8% of the total soluble leaf protein from genes with the clpP and atpB 5′-UTR, respectively. Interestingly, expression of NPTII from the promoter with the clpP 5′-UTR (0.26% NPTII) caused a mutant (chlorotic) phenotype, whereas plants accumulating approximately 0.8% NPTII from the atpB 5′-UTR were normal green, indicating that the mutant phenotype was independent of NPTII accumulation. Low levels of monocistronic clpP mRNA and accumulation of intron-containing clpP transcripts in the chlorotic leaves suggest competition between the clpP 5′-UTR in the chimeric transcript and the native clpP pre-mRNA (ratio 16:1) for an mRNA maturation factor. Because maturation of 11 other intron-containing mRNAs was unaffected in the chlorotic leaves, it appears that the factor is clpP specific. The mutant phenotype is correlated with reduced levels (approximately 2 times) of the ClpP1 protease subunit, supporting an important role for ClpP1 in chloroplast development.

Original languageEnglish
Pages (from-to)1600-1606
Number of pages7
JournalPlant Physiology
Volume129
Issue number4
DOIs
Publication statusPublished - Aug 2 2002
Externally publishedYes

Fingerprint

5' Untranslated Regions
5' untranslated regions
Chloroplasts
Tobacco
tobacco
chloroplasts
RNA
Phenotype
phenotype
mutants
Messenger RNA
Plastids
Introns
plastids
introns
promoter regions
Kanamycin Kinase
Untranslated Regions
leaves
kanamycin kinase

ASJC Scopus subject areas

  • Plant Science

Cite this

@article{6a6e76ad68b7437a8a94dc3c8e6ccff0,
title = "Overexpression of the clpP 5′-untranslated region in a chimeric context causes a mutant phenotype, suggesting competition for a clpP-specific RNA maturation factor in tobacco chloroplasts",
abstract = "The plastid ribosomal RNA (rrn) operon promoter was fused with DNA segments encoding the leader sequence (5′untranslated region [UTR]) of plastid mRNAs to compare their efficiency in mediating translation of a bacterial protein neomycin phosphotransferase (NPTII) in tobacco (Nicotiana tabacum) chloroplasts. In young leaves, NPTII accumulated at 0.26{\%} and 0.8{\%} of the total soluble leaf protein from genes with the clpP and atpB 5′-UTR, respectively. Interestingly, expression of NPTII from the promoter with the clpP 5′-UTR (0.26{\%} NPTII) caused a mutant (chlorotic) phenotype, whereas plants accumulating approximately 0.8{\%} NPTII from the atpB 5′-UTR were normal green, indicating that the mutant phenotype was independent of NPTII accumulation. Low levels of monocistronic clpP mRNA and accumulation of intron-containing clpP transcripts in the chlorotic leaves suggest competition between the clpP 5′-UTR in the chimeric transcript and the native clpP pre-mRNA (ratio 16:1) for an mRNA maturation factor. Because maturation of 11 other intron-containing mRNAs was unaffected in the chlorotic leaves, it appears that the factor is clpP specific. The mutant phenotype is correlated with reduced levels (approximately 2 times) of the ClpP1 protease subunit, supporting an important role for ClpP1 in chloroplast development.",
author = "Hiroshi Kuroda and Pal Maliga",
year = "2002",
month = "8",
day = "2",
doi = "10.1104/pp.004986",
language = "English",
volume = "129",
pages = "1600--1606",
journal = "Plant Physiology",
issn = "0032-0889",
publisher = "American Society of Plant Biologists",
number = "4",

}

TY - JOUR

T1 - Overexpression of the clpP 5′-untranslated region in a chimeric context causes a mutant phenotype, suggesting competition for a clpP-specific RNA maturation factor in tobacco chloroplasts

AU - Kuroda, Hiroshi

AU - Maliga, Pal

PY - 2002/8/2

Y1 - 2002/8/2

N2 - The plastid ribosomal RNA (rrn) operon promoter was fused with DNA segments encoding the leader sequence (5′untranslated region [UTR]) of plastid mRNAs to compare their efficiency in mediating translation of a bacterial protein neomycin phosphotransferase (NPTII) in tobacco (Nicotiana tabacum) chloroplasts. In young leaves, NPTII accumulated at 0.26% and 0.8% of the total soluble leaf protein from genes with the clpP and atpB 5′-UTR, respectively. Interestingly, expression of NPTII from the promoter with the clpP 5′-UTR (0.26% NPTII) caused a mutant (chlorotic) phenotype, whereas plants accumulating approximately 0.8% NPTII from the atpB 5′-UTR were normal green, indicating that the mutant phenotype was independent of NPTII accumulation. Low levels of monocistronic clpP mRNA and accumulation of intron-containing clpP transcripts in the chlorotic leaves suggest competition between the clpP 5′-UTR in the chimeric transcript and the native clpP pre-mRNA (ratio 16:1) for an mRNA maturation factor. Because maturation of 11 other intron-containing mRNAs was unaffected in the chlorotic leaves, it appears that the factor is clpP specific. The mutant phenotype is correlated with reduced levels (approximately 2 times) of the ClpP1 protease subunit, supporting an important role for ClpP1 in chloroplast development.

AB - The plastid ribosomal RNA (rrn) operon promoter was fused with DNA segments encoding the leader sequence (5′untranslated region [UTR]) of plastid mRNAs to compare their efficiency in mediating translation of a bacterial protein neomycin phosphotransferase (NPTII) in tobacco (Nicotiana tabacum) chloroplasts. In young leaves, NPTII accumulated at 0.26% and 0.8% of the total soluble leaf protein from genes with the clpP and atpB 5′-UTR, respectively. Interestingly, expression of NPTII from the promoter with the clpP 5′-UTR (0.26% NPTII) caused a mutant (chlorotic) phenotype, whereas plants accumulating approximately 0.8% NPTII from the atpB 5′-UTR were normal green, indicating that the mutant phenotype was independent of NPTII accumulation. Low levels of monocistronic clpP mRNA and accumulation of intron-containing clpP transcripts in the chlorotic leaves suggest competition between the clpP 5′-UTR in the chimeric transcript and the native clpP pre-mRNA (ratio 16:1) for an mRNA maturation factor. Because maturation of 11 other intron-containing mRNAs was unaffected in the chlorotic leaves, it appears that the factor is clpP specific. The mutant phenotype is correlated with reduced levels (approximately 2 times) of the ClpP1 protease subunit, supporting an important role for ClpP1 in chloroplast development.

UR - http://www.scopus.com/inward/record.url?scp=0037008190&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037008190&partnerID=8YFLogxK

U2 - 10.1104/pp.004986

DO - 10.1104/pp.004986

M3 - Article

C2 - 12177472

AN - SCOPUS:0037008190

VL - 129

SP - 1600

EP - 1606

JO - Plant Physiology

JF - Plant Physiology

SN - 0032-0889

IS - 4

ER -