An Overexpression Screen in Saccharomyces cerevisiae Identifies Novel Genes that Affect Endocytic Protein Trafficking
Version of Record online: 18 AUG 2011
© 2011 John Wiley & Sons A/S
Volume 12, Issue 11, pages 1592–1603, November 2011
How to Cite
Arlt, H., Perz, A. and Ungermann, C. (2011), An Overexpression Screen in Saccharomyces cerevisiae Identifies Novel Genes that Affect Endocytic Protein Trafficking. Traffic, 12: 1592–1603. doi: 10.1111/j.1600-0854.2011.01252.x
- Issue online: 6 OCT 2011
- Version of Record online: 18 AUG 2011
- Accepted manuscript online: 20 JUL 2011 11:52AM EST
- Received 28 January 2011, revised and accepted for publication 19 July 2011, uncorrected manuscript published online 20 July 2011, published online 18 August 2011
Additional Supporting Information may be found in the online version of this article:
Figure S1: CPY secretion upon gene overexpression. Cells carrying overexpression plasmids encoding the indicated genes were spotted onto SGC-URA plates and CPY secretion was assayed as described in Materials and Methods. Cells that strongly secrete the CPY-invertase chimera turn black and cells that secrete less than wild-type remain white. PET10 was excluded from further analyses.
Figure S2: Canavanine sensitivity upon gene overexpression. Serial dilutions of cells carrying overexpression plasmids encoding the indicated genes were spotted onto SGC-URA-ARG plates containing indicated canavanine concentrations. Plates were scanned after 4 days of growth at 30°C.
Figure S3: Phenotypic characteristics of uncharacterized gene deletions. A) Indicated yeast strains carrying CEN plasmids with indicated markers were grown in synthetic medium, followed by fluorescence microscopy. Size bar = 5 µm. B) Functionality of Cvt pathway upon gene deletion. Indicated yeast strains were grown to logarithmic phase, equal amounts of cells were harvested and lysed by addition of 10% trichloroacetic acid. Maturation of Ape1 was analyzed by western blot using an anti-Ape1 antibody.
Table S1: Yeast strains and plasmids used in this study.
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