In vivo bipartite interaction between the Hsp40 Sis1 and Hsp70 in Saccharomyces cerevisiae

Rebecca Aron; Nelson Lopez; William Walter; Elizabeth A Craig; Jill Johnson

doi:10.1534/genetics.104.037242

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In vivo bipartite interaction between the Hsp40 Sis1 and Hsp70 in Saccharomyces cerevisiae

Journal article

Open access

Peer reviewed

In vivo bipartite interaction between the Hsp40 Sis1 and Hsp70 in Saccharomyces cerevisiae

Rebecca Aron, Nelson Lopez, William Walter, Elizabeth A Craig and Jill Johnson

Genetics (Austin), Vol.169(4), pp.1873-1882

04/2005

DOI: https://doi.org/10.1534/genetics.104.037242

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https://hdl.handle.net/2376/116601

PMCID: PMC1449600

PMID: 15687271

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Abstract

Fungal Proteins - chemistry

Temperature

Cell Proliferation

Luciferases - metabolism

Molecular Sequence Data

Substrate Specificity

Immunoblotting

HSP40 Heat-Shock Proteins

Dose-Response Relationship, Drug

Saccharomyces cerevisiae - metabolism

Time Factors

Gene Deletion

Genes, Fungal

Binding, Competitive

Gene Expression Regulation, Fungal

Protein Structure, Tertiary

Amino Acid Sequence

Green Fluorescent Proteins - metabolism

Prions - metabolism

Enzyme-Linked Immunosorbent Assay

Gene Library

Protein Structure, Secondary

Genetic Techniques

Plasmids - metabolism

Protein Binding

HSP70 Heat-Shock Proteins - physiology

Protein Conformation

Saccharomyces cerevisiae Proteins - physiology

Mutation

Heat-Shock Proteins - physiology

Adenosine Triphosphatases - physiology

Saccharomyces cerevisiae Proteins - chemistry

The essential Hsp40, Sis1, is a J-protein cochaperone for the Ssa class of Hsp70's of Saccharomyces cerevisiae. Sis1 is required for the maintenance of the prion [RNQ(+)], as Sis1 lacking its 55-amino-acid glycine-rich region (G/F) does not maintain [RNQ(+)]. We report that overexpression of Sis1DeltaG/F in an otherwise wild-type strain had a negative effect on both cell growth and [RNQ(+)] maintenance, while overexpression of wild-type Sis1 did not. Overexpression of the related Hsp40 Ydj1 lacking its G/F region did not cause inhibition of growth, indicating that this dominant effect of Sis1DeltaG/F is not a characteristic shared by all Hsp40's. Analysis of small deletions within the SIS1 G/F region indicated that the observed dominant effects were caused by the absence of sequences known to be important for Sis1's unique cellular functions. These inhibitory effects of Sis1DeltaG/F were obviated by alterations in the N-terminal J-domain of Sis1 that affect interaction with Ssa's ATPase domain. In addition, a genetic screen designed to isolate additional mutations that relieved these inhibitory effects identified two residues in Sis1's carboxy-terminal domain. These alterations disrupted the interaction of Sis1 with the 10-kD carboxy-terminal regulatory domain of Ssa1, indicating that Sis1 has a bipartite interaction with Ssa in vivo.

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Title: In vivo bipartite interaction between the Hsp40 Sis1 and Hsp70 in Saccharomyces cerevisiae
Creators: Rebecca Aron - Department of Biochemistry, University of Wisconsin, Madison, 53706, USA
Nelson Lopez
William Walter
Elizabeth A Craig
Jill Johnson
Publication Details: Genetics (Austin), Vol.169(4), pp.1873-1882
Academic Unit: UNKNOWN
Publisher: United States
Grant note: R01 GM031107 / NIGMS NIH HHS GM31107 / NIGMS NIH HHS F31 GM018507 / NIGMS NIH HHS 5F31 GM18507-05 / NIGMS NIH HHS R37 GM031107 / NIGMS NIH HHS
Identifiers: 99900547574101842
Language: English
Resource Type: Journal article