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ERj1p has a basic role in protein biogenesis at the endoplasmic reticulum

Nature Structural & Molecular Biology volume 12, pages 1008–1014 (2005)Cite this article

Abstract

ERj1p is a membrane protein of the endoplasmic reticulum (ER) that can recruit the ER lumenal chaperone BiP to translating ribosomes. ERj1p can also modulate protein synthesis at initiation and is predicted to be a membrane-tethered transcription factor. Here we attribute the various functions of ERj1p to distinct regions within its cytosolic domain. A highly positively charged nonapeptide within this domain is necessary and sufficient for binding to ribosomes. Binding of ERj1p to ribosomes involves the 28S ribosomal RNA and occurs at the tunnel exit. Additionally, ERj1p has a dual regulatory role in gene expression: ERj1p inhibits translation in the absence of BiP, and another charged oligopeptide within the cytosolic domain of ERj1p mediates binding of the nuclear import factor importin β and import into the nucleus, thereby paving the way for subsequent action on genomic DNA.

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Figure 1: In vitro protein synthesis and ribosome binding of ERj1C are affected by the same N-terminal deletions.
Figure 2: Effects of different derivatives of WT 17-mer on in vitro protein synthesis and ribosome binding of ERj1C-ΔC85.
Figure 3: ERj1p modulates translation at the level of initiation.
Figure 4: The inhibitory effect of ERj1C-ΔC85 (ΔC85) on in vitro protein synthesis is prevented by the 28S rRNA of the 60S ribosomal subunit.
Figure 5: Binding of ERj1p to ribosomes occurs near the tunnel exit.
Figure 6: The inhibitory effect of ERj1p on in vitro protein synthesis is modulated by BiP.
Figure 7: Binding of ERj1C to nuclear import factors is affected by C-terminal deletions.
Figure 8: The role of ERj1p in protein synthesis at the ER surface.

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Acknowledgements

The plasmid that encodes GST-fusion mouse importin α and β was kindly provided by K. Weis (University of California, Berkeley, California, USA); yeast Ran was kindly donated by G. Schlenstedt (Universität des Saarlaudes, Homburg, Germany). We thank M. Albrecht and C. Völzing for help with bioinformatic analyses, to E. Herrmann for help with the statistical analysis and M. Lerner, S. Oberhauser and P. Scholtes for technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft, the Universität des Saarlandes and the Fonds der Chemischen Industrie.

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Authors and Affiliations

  1. Medizinische Biochemie und Molekularbiologie, Universität des Saarlandes, Homburg, D-66421, Germany

    Johanna Dudek, Markus Greiner, Anika Müller, Katharina Kopsch, Wolfgang Nastainczyk & Richard Zimmermann

  2. Department of Tumor Cell Biology, St. Jude Children's Hospital, Memphis, 38105, Tennessee, USA

    Linda M Hendershot

  3. Department of Molecular Sciences, University of Tennessee, Memphis, 38163, Tennessee, USA

    Linda M Hendershot

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  1. Johanna Dudek
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Correspondence to Richard Zimmermann.

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Supplementary information

Supplementary Fig. 1

In vitro protein synthesis is affected by different derivatives of WT 17-mer ERj1p to different extents. (PDF 127 kb)

Supplementary Fig. 2

The ribosome binding of ERj1C-ΔC85 and ERj1p, respectively, is not affected by BiP and BiP-R197H, respectively. (PDF 41 kb)

Supplementary Table 1

The oligopeptide motifs that are responsible for arrest of translation and nuclear import are distinct (PDF 4 kb)

Supplementary Table 2

The translational arrest activity of ERj1p but not the ribosome binding activity is suppressed by BiP: role of the J-domain (PDF 4 kb)

Supplementary Table 3

Highly charged oligopeptides that are present in various ligands of ribosomes (PDF 6 kb)

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Dudek, J., Greiner, M., Müller, A. et al. ERj1p has a basic role in protein biogenesis at the endoplasmic reticulum. Nat Struct Mol Biol 12, 1008–1014 (2005). https://doi.org/10.1038/nsmb1007

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