First published online February 25, 2005; 10.1104/pp.104.053637
Plant Physiology 137:848-862 (2005)
© 2005 American Society of Plant Biologists
BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES
Proteomic Characterization of Evolutionarily Conserved and Variable Proteins of Arabidopsis Cytosolic Ribosomes1,[w]
Ing-Feng Chang2,
Kathleen Szick-Miranda2,
Songqin Pan and
Julia Bailey-Serres*
Center for Plant Cell Biology, Department of Botany and Plant Sciences, University of California, Riverside, California 925210124 (I.-F.C., S.P., J.B.-S.); and Department of Biology, California State University, Bakersfield, California 93311 (K.S.-M.)
Analysis of 80S ribosomes of Arabidopsis (Arabidopsis thaliana) by use of high-speed centrifugation, sucrose gradient fractionation, one- and two-dimensional gel electrophoresis, liquid chromatography purification, and mass spectrometry (matrix-assisted laser desorption/ionization time-of-flight and electrospray ionization) identified 74 ribosomal proteins (r-proteins), of which 73 are orthologs of rat r-proteins and one is the plant-specific r-protein P3. Thirty small (40S) subunit and 44 large (60S) subunit r-proteins were confirmed. In addition, an ortholog of the mammalian receptor for activated protein kinase C, a tryptophan-aspartic acid-domain repeat protein, was found to be associated with the 40S subunit and polysomes. Based on the prediction that each r-protein is present in a single copy, the mass of the Arabidopsis 80S ribosome was estimated as 3.2 MD (1,159 kD 40S; 2,010 kD 60S), with the 4 single-copy rRNAs (18S, 26S, 5.8S, and 5S) contributing 53% of the mass. Despite strong evolutionary conservation in r-protein composition among eukaryotes, Arabidopsis 80S ribosomes are variable in composition due to distinctions in mass or charge of approximately 25% of the r-proteins. This is a consequence of amino acid sequence divergence within r-protein gene families and posttranslational modification of individual r-proteins (e.g. amino-terminal acetylation, phosphorylation). For example, distinct types of r-proteins S15a and P2 accumulate in ribosomes due to evolutionarily divergence of r-protein genes. Ribosome variation is also due to amino acid sequence divergence and differential phosphorylation of the carboxy terminus of r-protein S6. The role of ribosome heterogeneity in differential mRNA translation is discussed.
1 This research was supported by the National Science Foundation (grant no. DBI 021187 to J.B.-S.), by the U.S. Department of Agriculture (grant no. 00353019108 to J.B.-S.), and by the Ministry of Education, Republic of China, Taiwan (grant to I.-F.C.).
2 These authors contributed equally to the paper.
[w] The online version of this article contains Web-only data.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.053637.
* Corresponding author; e-mail serres{at}ucr.edu; fax 9518274437.
Received September 16, 2004;
returned for revision November 16, 2004;
accepted November 23, 2004.
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