Distribution of membrane-bound and free ribosomes in growing yeast

Schneider, Elisabeth; Lochmann, Ernst-Randolf; Lother, Heinz

doi:10.1016/0005-2787(76)90044-7

Cited by 13 publications

(6 citation statements)

References 14 publications

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“…This result suggests either that there are diffusional barriers for particles of this size or that the ribosomes are attached to large structures that diffuse slowly. Nevertheless, most ribosomes have been found free in the cytoplasm and not bound to endoplasmic reticulum in yeast (39). It is possible, that ribosomes are actually loosely associated with either membranes or other slowly moving cellular structures and are released on cell disruption.…”

Section: Discussionmentioning

confidence: 99%

Yeast Ribosomal Stalk Heterogeneity In Vivo Shown by Two-Photon FCS and Molecular Brightness Analysis

García-Marcos

Sánchez

Parada

et al. 2008

Biophysical Journal

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The stalk of Saccharomyces cerevisiae ribosomes contains, on average, five distinct proteins, namely P0 and four acidic proteins, P1alpha, P1beta, P2alpha, and P2beta. Each ribosome contains only one copy of P0, but the distribution of the acidic proteins among the ribosome population in vivo has not been determined. Using two-photon fluorescence correlation spectroscopy and scanning FCS, on cells expressing EGFP-tagged P0, P1, and P2 proteins, we show, with brightness analysis, that individual yeast ribosomes in vivo are compositionally heterogeneous in regard to P1alpha, P1beta, P2alpha, and P2beta. These results are relevant to the hypothesis, based on in vitro studies, that the overall cellular pattern of expressed proteins can be determined by the distribution of the stalk proteins among the ribosome population.

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Section: Discussionmentioning

confidence: 99%

Yeast Ribosomal Stalk Heterogeneity In Vivo Shown by Two-Photon FCS and Molecular Brightness Analysis

García-Marcos

Sánchez

Parada

et al. 2008

Biophysical Journal

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“…To test whether the processed chains were still attached to ribosomes, we isolated the RNCs from a total import reaction by solubilizing the mitochondria in 1% Triton X-100 and sedimentation through a sucrose cushion ( Figure 1B). Triton X-100 solubilizes membranes but leaves ribosomes intact (Schneider et al, 1976). The majority of both precursor and mature Mdh1p was recovered in the ribosomal pellet (R-Pel), indicating that they were still attached to ribosomes.…”

Section: Nac Stimulates Mitochondrial Protein Importmentioning

confidence: 95%

Nascent Polypeptide–associated Complex Stimulates Protein Import into Yeast Mitochondria

Fünfschilling

Rospert

1999

MBoC

148

127

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To identify yeast cytosolic proteins that mediate targeting of precursor proteins to mitochondria, we developed an in vitro import system consisting of purified yeast mitochondria and a radiolabeled mitochondrial precursor protein whose C terminus was still attached to the ribosome. In this system, the N terminus of the nascent chain was translocated across both mitochondrial membranes, generating a translocation intermediate spanning both membranes. The nascent chain could then be completely chased into the mitochondrial matrix after release from the ribosome. Generation of this import intermediate was dependent on a mitochondrial membrane potential, mitochondrial surface proteins, and was stimulated by proteins that could be released from the ribosomes by high salt. The major salt-released stimulatory factor was yeast nascent polypeptide–associated complex (NAC). Purified NAC fully restored import of salt-washed ribosome-bound nascent chains by enhancing productive binding of the chains to mitochondria. We propose that ribosome-associated NAC facilitates recognition of nascent precursor chains by the mitochondrial import machinery.

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“…The cell free extracts were prepared according to the described procedure (JAKUBOWICZ et al 1993). Membrane free 80 S ribosomes were released from the endoplasmic reticulum by 1 % Triton X-100 treatment of rnicrosomal fraction (SCHNEIDER et al 1976). The concentration of ribosomes (1 1 OD,, = I mg/ml) was estimated according…”

Section: Methodsmentioning

confidence: 99%

The acidic ribosomal proteins of different yeast species. Phosphorylation by ribosome‐associated protein kinases

Cytryńska

Wojda

Jakubowicz

1995

J. Basic Microbiol.

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Two major ribosomal proteins of Mr 13 kDa and 38 kDa were identified as phosphorylation substrates for ribosome-bound protein kinases from different yeast species. The phosphorylation level was much higher in ribosomes from the cells collected at the exponential growth phase, than in those from the stationary phase. Isoelectrofocusing of the protein band of 13 kDa and subsequent silver staining allowed to identify from four in the case of Pichia stipitis up to ten in Saccharomyces cerevisiae. Saccharomycodes Ludwigii, Torulopsis utilis and Kloeckera apiculata individual peptides. In the most of the yeast species studies five phosphorylated peptides were observed. However, only one or two such phosphopeptides were detected in Pichia stipitis and Trichosporon cutaneum ribosomes, respectively. The same phosphoprotein forms were identified in the in vivo 32P-labelling experiments.

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Distribution of membrane-bound and free ribosomes in growing yeast

Cited by 13 publications

References 14 publications

Yeast Ribosomal Stalk Heterogeneity In Vivo Shown by Two-Photon FCS and Molecular Brightness Analysis

Yeast Ribosomal Stalk Heterogeneity In Vivo Shown by Two-Photon FCS and Molecular Brightness Analysis

Nascent Polypeptide–associated Complex Stimulates Protein Import into Yeast Mitochondria

The acidic ribosomal proteins of different yeast species. Phosphorylation by ribosome‐associated protein kinases

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