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Functional studies of vesicular transport in yeast

Barmark, Gunilla (2005). Functional studies of vesicular transport in yeast. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., Acta Universitatis agriculturae Sueciae, 1652-6880 ; 2005:110
ISBN 91-576-6909-0
[Doctoral thesis]

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Abstract

A basic feature of eukaryotic cells is their compartmental organisation. To maintain this cellular order, a strictly regulated intracellular transport is essential. The compartments are defined by their lipid membranes and the transport between different compartments is therefore mediated by lipid vesicles. The yeast Saccharomyces cerevisiae is a very suitable model organism for studies of intracellular transport. In order to find new yeast genes involved in intracellular transport, a screen was performed using a knockout yeast strain collection and two drugs known to interfere with intracellular transport, monensin and brefeldin A. In an initial screen, seven new genes were found (MON1-2 and BRE1-5) that caused an increased sensitivity to either drug when knocked out. When the monensin screen was extended to the entire yeast genome, a total of 63 sensitive strains were found. Most of the encoded proteins are known to be involved in post-Golgi transport. Some of the 63 monensin-sensitive knockout mutants showed synthetic interactions with a vma1 knockout mutant, ranging from complete synthetic lethality to reduced growth. These interactions were specific for genes involved in retrograde transport. One of the new genes found in the screen, MON1, is involved in transport to the vacuole. A mon1 deletion strain is impaired in maturation of proaminopeptidase both during starvation and during nutrient-rich conditions, indicating that Mon1p is involved in both the autophagic and the cytoplasm-to-vacuole targeting pathways. A GFP-Mon1p fusion protein localizes to the cytosol, and to punctate structures in the cytosol. Furthermore, the autophagosomal marker protein GFP-Aut7p accumulates in mon1 deleted cells at punctate structures near the vacuole. The two syntaxins, Sso1p and Sso2p, are involved in the fusion of vesicles to the plasma membrane, where they can functionally replace each other. However, the SSO1 gene also has an essential function in meiosis, which SSO2 cannot provide. Gene fusions between SSO1 and SSO2 were used to map this meiosis-specific function to two short elements within the 3'-untranslated region of the SSO1 gene.

Authors/Creators:Barmark, Gunilla
Title:Functional studies of vesicular transport in yeast
Year of publishing :November 2005
Volume:2005:110
Number of Pages:46
Papers/manuscripts:
NumberReferences
I.Murén E., Öyen M., Barmark G., Ronne H., (2001) Identification of yeast deletion strains that are hypersensitive to brefeldin A or monensin, two drugs that affect intracellular transport. Yeast 18:163-172.
II.Meiling-Wesse K., Barth H., Voss C., Barmark G., Murén E., Ronne H., Thumm M., (2002) Yeast Mon1p/Aut12p functions in vacuolar fusion of autophagosomes and cvt-vesicles. FEBS Letters 530: 174-180.
III.Barmark G., Gustavsson M., Larsson J., Murén E., Ronne H., Functional genomics of monensin sensitivity in yeast: implications for post-Golgi traffic. Manuscript.
IV. Barmark G., Murén E., Harnemark H., Gustavsson M., Ronne H. Mapping of sequences in the 3’-untranslated region of the yeast plasma membrane syntaxin gene SSO1 that are required for sporulation. Manuscript.
Place of Publication:Uppsala
Publisher:Dept. of Plant Biology and Forest Genetics, Swedish University of Agricultural Sciences
ISBN for printed version:91-576-6909-0
ISSN:1652-6880
Language:English
Publication Type:Doctoral thesis
Full Text Status:Public
Agris subject categories.:F Plant production > F30 Plant genetics and breeding
Subjects:Not in use, please see Agris categories
Agrovoc terms:monensin, proteins, saccharomyces cerevisiae, eucaryotic cells, vacuoles, yeasts, genes
Keywords:Brefeldin A, monensin, Saccharomyces cerevisiae, syntaxin, vacuole, vesicular transport
URN:NBN:urn:nbn:se:slu:epsilon-805
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-805
ID Code:979
Department:(NL, NJ) > Dept. of Plant Biology and Forest Genetics (until 131231)
Deposited By: Gunilla Barmark
Deposited On:15 Nov 2005 00:00
Metadata Last Modified:02 Dec 2014 10:08

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