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Zerges, W.

Publications and source records attributed to Zerges, W..

3 recordsLinked to original sources

Membranous translation platforms in the chloroplast of Chlamydomonas reinhardtii

A small genome in chloroplasts encodes polypeptide subunits of the photosynthetic electron transport complexes in the membranes of thylakoid vesicles in the chloroplast stroma. Trans-membrane subunits of these complexes undergo co-translational membrane insertion during their synthesis by ribosomes of the bacterial-like genetic system of this semiautonomous organelle. While thylakoid membranes are sites of translation, evidence in the unicellular alga Chlamydomonas reinhardtii supports translation also on non-canonical membranes in a discrete translation-zone in the chloroplast. To characterize the membranous platforms for translation and the biogenesis of thylakoid membrane, we profiled membranes during chloroplast development, using the yellow-in-the-dark 1 mutant, and carried out proteomic analyses on membranes of interest. The results support roles of two membrane types in preliminary and ongoing stages of translation: a "low-density membrane" and a denser "chloroplast translation membrane", respectively. These roles are based on correlations of the relative levels of each membrane type and the translational status of the chloroplast before, during and after chloroplast differentiation and results of proteomic analyses. Our results support a model of photosynthesis complex biogenesis in a spatiotemporal "assembly line" involving LDM and CTM as sequential stages leading to photosynthetic thylakoid membranes.

cell biology↗

Chloroplast-localized translation for protein targeting in Chlamydomonas reinhardtii

Translation is localized within cells to target proteins to their proper locations. We asked whether translation occurs on the chloroplast surface in Chlamydomonas and, if so, whether it is involved in co-translational protein targeting, aligned spatially with localized translation by the bacterial-type ribosomes within this organelle, or both. Our results reveal a domain of the chloroplast envelope which is bound by translating ribosomes. Purified chloroplasts retained ribosomes and mRNAs encoding two chloroplast proteins specifically on this "translation domain", but not a mRNA encoding a cytoplasmic protein. Ribosomes clusters were seen on this domain by electron tomography. Activity of the chloroplast-bound ribosomes is supported by results of the ribopuromycylation and puromycin-release assays. Co-translational chloroplast protein import is supported by nascent polypeptide dependency of the ribosome-chloroplast associations. This cytoplasmic translation domain aligns localized translation by organellar bacterial-type ribosomes in the chloroplast. This juxtaposition the dual translation systems facilitates the targeting and assembly of the polypeptide products. One-Sentence SummaryTranslation is localized to a domain of the chloroplast envelope for co-translational protein targeting in Chlamydomonas.

cell biology↗

Oxidized RNA Bodies compartmentalize translation quality control in Saccharomyces cerevisiae

Cytoplasmic RNA granules compartmentalize phases of the translation cycle. We previously reported on the localization of oxidized RNA in human cells to cytoplasmic foci called oxidized RNA bodies (ORBs). Oxidized mRNAs are substrates of translation quality control, wherein defective mRNAs and nascent polypeptides are released from stalled ribosomes and targeted for degradation. Therefore, we asked whether ORBs compartmentalize translation quality control. Here, we identify ORBs in Saccharomyces cerevisiae and characterize them using fluorescence microscopy and proteomics. ORBs are RNA granules that are distinct from processing bodies and stress granules. Several lines of evidence support a role of ORBs in the compartmentalization of central steps in the translation quality control pathways No-Go mRNA decay and ribosome quality control. Active translation is required by both translation quality control and ORBs. ORBs contain two substrates of translation quality control: oxidized RNA and a stalled mRNA-ribosome-nascent chain complex. Translation quality control factors localize to ORBs. Translation quality control mutants have altered ORB number per cell, size, or both. Therefore, ORBs are an intracellular hub of translational quality control.

cell biology↗