Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells

Samuel B Stephens; Rebecca D Dodd; Rachel S Lerner; Brook M Pyhtila; Christopher V Nicchitta

doi:10.1007/978-1-59745-033-1_14

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Journal article

Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells

Samuel B Stephens, Rebecca D Dodd, Rachel S Lerner, Brook M Pyhtila and Christopher V Nicchitta

Methods in molecular biology (Clifton, N.J.), Vol.419, pp.197-214

2008

DOI: 10.1007/978-1-59745-033-1_14

PMID: 18369985

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Abstract

All eukaryotic cells display a dramatic partitioning of mRNAs between the cytosol and endoplasmic reticulum (ER) compartments-mRNAs encoding secretory and integral membrane proteins are highly enriched on ER-bound ribosomes and mRNAs encoding cytoplasmic/nucleoplasmic proteins are enriched on cytosolic ribosomes. In current views, this partitioning phenomenon occurs through positive selection-mRNAs encoding signal sequence-bearing proteins are directed into the signal recognition particle pathway early in translation and trafficked as mRNA/ribosome/nascent polypeptide chain complexes to the ER. In the absence of an encoded signal sequence, mRNAs undergo continued translation on cytosolic ribosomes. Recent genome-wide analyses of mRNA partitioning between the cytosol and the ER compartments have identified subsets of mRNAs that are non-canonically partitioned to the ER-although lacking an encoded signal sequence, they are translated on ER-bound ribosomes. These findings suggest that multiple, and as yet unidentified, pathways exist for directing mRNA partitioning in the cell. In this contribution, we briefly review the literature describing the subcellular partitioning patterns of mRNAs and present a detailed methodology for studying this fundamental, yet poorly understood process.

NIH 3T3 Cells

Oligonucleotide Probes

RNA, Messenger - isolation & purification

Jurkat Cells

Humans

RNA, Messenger - genetics

Cercopithecus aethiops

Endoplasmic Reticulum - metabolism

Ribosomes - metabolism

Centrifugation, Isopycnic

Detergents

RNA, Messenger - metabolism

Cell Compartmentation

Blotting, Northern

Animals

Electrophoresis, Agar Gel

Cytosol - metabolism

Mice

HeLa Cells

COS Cells

Details

Title: Subtitle: Analysis of mRNA partitioning between the cytosol and endoplasmic reticulum compartments of mammalian cells
Creators: Samuel B Stephens - Department of Cell Biology, Duke University Medical Center, Durham, NC, USA
Rebecca D Dodd
Rachel S Lerner
Brook M Pyhtila
Christopher V Nicchitta
Resource Type: Journal article
Publication Details: Methods in molecular biology (Clifton, N.J.), Vol.419, pp.197-214
DOI: 10.1007/978-1-59745-033-1_14
PMID: 18369985
eISBN: 9781597450331; 1597450332
ISSN: 1064-3745
eISSN: 1940-6029
Grant note: GM077382 / NIGMS NIH HHS
Language: English
Date published: 2008
Academic Unit: Hematology, Oncology, and Blood & Marrow Transplantation; Fraternal Order of Eagles Diabetes Research Center; Endocrinology and Metabolism; Internal Medicine
Record Identifier: 9984094524302771

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