Adenosine Triphosphatases; Genetics; Physiology; Saccharomyces cerevisiae; Ion Transport
Cellular & Molecular Physiology: Membrane Protein Sorting and Trafficking | Membrane Proteins - Pumps and Transporters
Research in Carolyn Slayman’s laboratory uses the plasma-membrane H+-ATPase of yeast (Saccharomyces cerevisiae) as a simple model for studies on P-type pumps, a physiologically important family that includes the Na+,K+-, H+,K+-, and Ca2+-ATPases of animal cells. These pumps control the ionic composition of cells; many of them also serve as important drug targets (e.g., for cardiac glycosides in the case of Na+,K+-ATPase and anti-ulcer drugs in the case of gastric H+,K+-ATPase).
Specialized Terms: Genetics of Ion Transport
Extensive Research Description
Over the past several years, the Slayman laboratory has constructed a large collection of site-directed mutants and used them to explore structure-function relationships in key regions of the 100 kDa H+-ATPase. Results of particular significance include: (1) discovery of a 13-amino acid stretch in stalk segment 4 within which mutations disrupt the interaction between the catalytic domain of the ATPase (located in the cytoplasm) and the proton channel (embedded in the membrane); and (2) clear implication of one face of stalk segment 5 in the metabolic activation of the ATPase by glucose. Cysteine residues have now been introduced into both regions, and methods are being developed to track conformational changes by means of fluorescent sulfhydryl reagents. With a recently published high resolution structure of sarcoplasmic reticulum Ca2+-ATPase as a template, biophysical data from the H+-ATPase mutants should yield helpful insights into the molecular mechanism by which ATP hydrolysis is coupled to cation transport.
In parallel, the H+-ATPase is being used as a tool to probe the way in which newly synthesized plasma membrane proteins are delivered from the endoplasmic reticulum to the cell surface, as well as the quality control mechanisms that remove poorly folded or otherwise defective proteins. During the past year, a nested series of truncations has been constructed and has provided intriguing evidence for an oligomerization step at the C-terminus of the ATPase; oligomerization appears to occur early in the secretory pathway and to be essential for normal biogenesis.
Tandem phosphorylation of Ser-911 and Thr-912 at the C-terminus of yeast plasma membrane H+-ATPase leads to glucose-dependent activation.
Lecchi, S., Nelson, C.J., Allen, K.E., Swaney, D.L., Thompson, K.L., Coon, J.J., Sussman, M.R., and Slayman, C.W. (2007) Tandem phosphorylation of Ser-911 and Thr-912 at the C-terminus of yeast plasma membrane H+-ATPase leads to glucose-dependent activation. J.Biol.Chem. 282:35471-35481. (PMID: 17932035)
Role of transmembrane segment M8 in the biogenesis and function of yeast plasma-membrane H+-ATPase.
Guerra, G., Petrov, V.V., Allen, K.E., Miranda, M., Pardo, J.P., and Slayman, C.W. (2007) Role of transmembrane segment M8 in the biogenesis and function of yeast plasma-membrane H+-ATPase. Biochim.Biophys.Acta 1768:2383-2392. (PMID: 17573037)
Effects of C-terminal truncations on trafficking of the yeast plasma membrane H+-ATPase.
Mason, A. B., Allen, K. E., and Slayman, C. W. (2006) Effects of C-terminal truncations on trafficking of the yeast plasma membrane H+-ATPase. Biological Chemistry 281:23887-23898
Conformational changes of yeast plasma membrane H+-ATPase during activation by glucose: role of Thr912 in the carboxy-terminal tail.
Lecchi, S., Allen, K. E., Pardo, J. P., Mason, A. B., and Slayman, C. W. (2005) Conformational changes of yeast plasma membrane H+-ATPase during activation by glucose: role of Thr912 in the carboxy-terminal tail. Biochemistry 44:16624-16632
Full List of PubMed Publications
- Goldgof GM, Durrant JD, Ottilie S, Vigil E, Allen KE, Gunawan F, Kostylev M, Henderson KA, Yang J, Schenken J, LaMonte GM, Manary MJ, Murao A, Nachon M, Stanhope R, Prescott M, McNamara CW, Slayman CW, Amaro RE, Suzuki Y, Winzeler EA: Comparative chemical genomics reveal that the spiroindolone antimalarial KAE609 (Cipargamin) is a P-type ATPase inhibitor. Sci Rep. 2016 Jun 13; 2016 Jun 13. PMID: 27291296
- Mason AB, Allen KE, Slayman CW: C-terminal truncations of the Saccharomyces cerevisiae PMA1 H+-ATPase have major impacts on protein conformation, trafficking, quality control, and function. Eukaryot Cell. 2014 Jan; 2013 Nov 1. PMID: 24186948
- Rivetta A, Allen KE, Slayman CW, Slayman CL: Coordination of K+ transporters in neurospora: TRK1 is scarce and constitutive, while HAK1 is abundant and highly regulated. Eukaryot Cell. 2013 May; 2013 Mar 8. PMID: 23475706
- Sherwin R, Slayman C, Rockwell S, Herold K, Williams K, Carson R, Mane S, Seow H, Max J, Johnson T: Yale Center for Clinical Investigation: leveraging industry partnerships and research cores. Clin Transl Sci. 2012 Dec. PMID: 23253663
- Lecchi S, Nelson CJ, Allen KE, Swaney DL, Thompson KL, Coon JJ, Sussman MR, Slayman CW: Tandem phosphorylation of Ser-911 and Thr-912 at the C terminus of yeast plasma membrane H+-ATPase leads to glucose-dependent activation. J Biol Chem. 2007 Dec 7; 2007 Oct 11. PMID: 17932035
- Mason AB, Allen KE, Slayman CW: Effects of C-terminal truncations on trafficking of the yeast plasma membrane H+-ATPase. J Biol Chem. 2006 Aug 18; 2006 Jun 2. PMID: 16751629
- Lecchi S, Allen KE, Pardo JP, Mason AB, Slayman CW: Conformational changes of yeast plasma membrane H(+)-ATPase during activation by glucose: role of threonine-912 in the carboxy-terminal tail. Biochemistry. 2005 Dec 20. PMID: 16342953
- Slayman CW, Miranda M, Pardo JP, Allen KE: Use of a fluorescent maleimide to probe structure-function relationships in stalk segments 4 and 5 of the yeast plasma-membrane H+-ATPase. Ann N Y Acad Sci. 2003 Apr. PMID: 12763792
- Miranda M, Pardo JP, Allen KE, Slayman CW: Stalk segment 5 of the yeast plasma membrane H(+)-ATPase. Labeling with a fluorescent maleimide reveals a conformational change during glucose activation. J Biol Chem. 2002 Oct 25; 2002 Aug 6. PMID: 12169695
- Ferreira T, Mason AB, Pypaert M, Allen KE, Slayman CW: Quality control in the yeast secretory pathway: a misfolded PMA1 H+-ATPase reveals two checkpoints. J Biol Chem. 2002 Jun 7; 2002 Feb 27. PMID: 11877403