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Titel: Quantitative Proteomics and Differential Protein Abundance Analysis after Depletion of Putative mRNA Receptors in the ER Membrane of Human Cells Identifies Novel Aspects of mRNA Targeting to the ER
VerfasserIn: Bhadra, Pratiti
Schorr, Stefan
Lerner, Monika
Nguyen, Duy
Dudek, Johanna
Förster, Friedrich
Helms, Volkhard
Lang, Sven
Zimmermann, Richard
Sprache: Englisch
Titel: Molecules
Bandnummer: 26
Heft: 12
Verlag/Plattform: MDPI
Erscheinungsjahr: 2021
Freie Schlagwörter: endoplasmic reticulum
mRNA targeting
protein targeting
protein import
membrane protein insertion
protein translocation
Sec61 complex
TIGER domain
label-free quantitative mass spectrometry
differential protein abundance analysis
DDC-Sachgruppe: 500 Naturwissenschaften
610 Medizin, Gesundheit
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: In human cells, one-third of all polypeptides enter the secretory pathway at the endoplasmic reticulum (ER). The specificity and efficiency of this process are guaranteed by targeting of mRNAs and/or polypeptides to the ER membrane. Cytosolic SRP and its receptor in the ER membrane facilitate the cotranslational targeting of most ribosome-nascent precursor polypeptide chain (RNC) complexes together with the respective mRNAs to the Sec61 complex in the ER membrane. Alternatively, fully synthesized precursor polypeptides are targeted to the ER membrane post-translationally by either the TRC, SND, or PEX19/3 pathway. Furthermore, there is targeting of mRNAs to the ER membrane, which does not involve SRP but involves mRNA- or RNC-binding proteins on the ER surface, such as RRBP1 or KTN1. Traditionally, the targeting reactions were studied in cell-free or cellular assays, which focus on a single precursor polypeptide and allow the conclusion of whether a certain precursor can use a certain pathway. Recently, cellular approaches such as proximity-based ribosome profiling or quantitative proteomics were employed to address the question of which precursors use certain pathways under physiological conditions. Here, we combined siRNA-mediated depletion of putative mRNA receptors in HeLa cells with label-free quantitative proteomics and differential protein abundance analysis to characterize RRBP1- or KTN1- involving precursors and to identify possible genetic interactions between the various targeting pathways. Furthermore, we discuss the possible implications on the so-called TIGER domains and critically discuss the pros and cons of this experimental approach.
DOI der Erstveröffentlichung: 10.3390/molecules26123591
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-342615
hdl:20.500.11880/31459
http://dx.doi.org/10.22028/D291-34261
ISSN: 1420-3049
Datum des Eintrags: 1-Jul-2021
Bezeichnung des in Beziehung stehenden Objekts: Supplementary Materials
In Beziehung stehendes Objekt: https://www.mdpi.com/1420-3049/26/12/3591/s1
Fakultät: M - Medizinische Fakultät
ZE - Zentrale Einrichtungen
Fachrichtung: M - Medizinische Biochemie und Molekularbiologie
ZE - Zentrum für Bioinformatik(ZBI)
Professur: M - Keiner Professur zugeordnet
ZE - Sonstige
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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