Cited by 60RNA Research and SplicingMitochondrial Function and PathologyRNA and protein synthesis mechanisms

The mitochondrial message-specific mRNA protectors Cbp1 and Pet309 are associated in a high-molecular weight complex

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Krause, Kirsten, Lopes de Souza, Renata, Roberts, Douglas G.W., Dieckmann, Carol L.·2004·Molecular Biology of the Cell

Abstract

In Saccharomyces cerevisiae, the nuclear-encoded protein Cbp1 promotes stability and translation of mitochondrial cytochrome b transcripts through interaction with the 5' untranslated region. Fusion of a biotin binding peptide tag to the C terminus of Cbp1 has now allowed detection in mitochondrial extracts by using peroxidase-coupled avidin. Cbp1 is associated with the mitochondrial membranes when high ionic strength extraction conditions are used. However, the protein is easily solubilized by omitting salt from the extraction buffer, which suggests Cbp1 is loosely associated with the membrane through weak hydrophobic interactions. Gel filtration analysis and blue native PAGE showed that Cbp1 is part of a single 900,000-Da complex. The complex was purified using the biotin tag and a sequence-specific protease cleavage site. In addition to Cbp1, the complex contains several polypeptides of molecular weights between 113 and 40 kDa. Among these, we identified another message-specific factor, Pet309, which promotes the stability and translation of mitochondrial cytochrome oxidase subunit I mRNA. A hypothesis is presented in which the Cbp1-Pet309 complex contains several message-specific RNA binding proteins and links transcription to translation of the mRNAs at the membrane.

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  1. Gharahdaghi, F., Weinberg, C.R., Meagher, D.A., Imai, B.S., and Mishe, S.M. (1999). Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity. Electrophoresis 20, 601-605. [DOI] [PubMed] [Google Scholar]

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Authors
Krause, K., Lopes de Souza, R., Roberts, D. G. W., & Dieckmann, C. L.
Affiliations
  • University of Arizona (US)
  • Department of Biochemistry and Molecular Biophysics, University of Arizona, Tucson, Arizona 85721
Venue
Molecular Biology of the Cell
Published
2004-06
Volume
15
Issue
6
Pages
2674-2683
ISSN
  • 1059-1524 (ISSN-L)
  • 1939-4586
OpenAlex
W2026484869
Cited by
60
References
62
Citations by year
2025: 2 · 2024: 1 · 2022: 1 · 2020: 1 · 2019: 3 · 2018: 2 · 2017: 3 · 2016: 1 · 2015: 2 · 2014: 1 · 2013: 3 · 2012: 6
FWCI
2.13
Type
journal article
Language
en
Open access
Locations
  • Molecular Biology of the Cell (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • Deutscher Akademischer Austauschdienst
  • National Institutes of Health
  • National Institute of General Medical Sciences (R01GM034893, GM34893)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
Keywords
CBP1, mitochondrial translation, 5' untranslated region, RNA-binding proteins, high molecular weight protein complexes, transcription-translation coupling, Saccharomyces cerevisiae
MeSH terms
RNA, Mitochondrial, Buffers, Chromatography, Gel, Intracellular Membranes, Membrane Proteins, Molecular Weight, Peptide Initiation Factors, Protein Binding, RNA, Saccharomyces cerevisiae, Salts, Solubility, Sonication, Substrate Specificity, RNA-Binding Proteins, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Mitochondrial Proteins, Saccharomyces cerevisiae Proteins, Multiprotein Complexes

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