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FACULTY

Michael A. McMurray


Associate Professor​

Michael McMurray, Ph.D.

Cell and Developmental Biology

Ph.D., University of Washington and Fred Hutchinson Cancer Research Center, 2004

Research Interests​

Mechanisms of assembly and inheritance of dynamic macromolecular structures: Higher-order septin assemblies in budding yeast

Cellular and molecular mechanisms of yeast gametogenesis

Contact

Office Location: RC-1 South, Room 12117

Mailing Address:

Mail Stop 8108​
12801 East 17th Avenue
Aurora, CO 80045

Phone: 303-724-6569
Fax: 303-724-3420
Email: Michael.McMurray@ucdenver.edu

Departmental Affiliations

Cell and Developmental Biology

Graduate Program Affiliations

Cell Biology, Stem Cells, and Development Program (CSDV)

Molecular Biology Program (MOLB)

Biomedical Sciences Training (BSP)

Medical Scientist Training Program (MSTP)​

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My research focuses on identifying molecular mechanisms underlying the assembly of macromolecular complexes, with a focus on multisubunit complexes formed by septin proteins. All cellular processes require the function of multisubunit complexes, and while much attention has been given to solving the final structures of such assemblies, comparatively little is known about how individual subunits adopt oligomerization-competent conformations and find their partner subunits in the crowded, dynamic cellular milieu. Below is a summary of the past research from our group.

  • We first used unbiased genetic screening to find evidence that guanine nucleotide binding by septin proteins plays an entirely structural role during filament assembly, which can be bypassed by specific mutations in a key septin-septin oligomerization interface. Considering that one of these mutations has been found to cause male infertility in humans, these findings have compelling clinical relevance.

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The structure of a homodimer of the human septin SEPT2 bound to the nonhydrolyzable GTP analog GppNHp (PDB 3FTQ) with the residues corresponding to those we found in temperature-sensitive yeast rendered as spheres. GppNHp is shown in orange. From Weems, et al GENETICS 2013.

  • We then investigated a phenomenon in which, when cells have a choice between a wild-type and a mutant version of a given septin, mutant septins unable to bind nucleotides are discriminated against during higher-order assembly, despite their ability to function normally under the conditions tested. We discovered that prolonged interaction between nascent mutant polypeptides and cytosolic chaperones results in a kinetic delay in the acquisition by the mutant septin of a conformation competent for oligomerization with other septins. Our findings challenged the dominant paradigm that interactions between chaperones and their clients always promote mutant protein function, and provided a new way of thinking about how chaperone interactions might influence the ability of mutant alleles of proteins to perturb the functions of the wild type.

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Model for chaperone-mediated quality control of higher-order septin assembly. Nascent septin polypeptides emerging from the ribosome encounter a number of cytosolic chaperones during subsequent de novo folding. Wild-type septins efficiently adopt quasi-native conformations, thereby burying hydrophobic residues and escaping chaperone-mediated sequestration. Heterodimerization with other septins—the first oligomerization step toward septin filament assembly—occurs concomitant with exit from the chamber of the cytosolic chaperonin CCT (also called TRiC). Mutant septins that inefficiently fold the G heterodimerization interface are slower to achieve a conformation allowing chaperone release. Interactions with the prefoldin complex (PFD), the Hsp40 chaperone Ydj1, and the disaggregase Hsp104 are particularly prolonged, leading to a delay in availability of the mutant septin for hetero-oligomerization. From Johnson, et al Mol Biol Cell 2015.

  • We went on to develop an entirely new technique for monitoring, for the first time in living cells, the step-wise pathway of assembly of protein complexes, and used it to show how nucleotide-induced conformational changes drive the progression of such an ordered pathway for yeast septins. Moreover, we generated and validated a new model for the role of nucleotide hydrolysis by septins during oligomerization, and found that slow GTPase activity by a key septin subunit drives the formation of distinct hetero-oligomers comprised of alternative subunits. Our “GTPase timer” model predicted that changes in the ratio of GTP to GDP in the cytosol would influence the subunit composition of septin complexes. Our experiments supported this model and further provided a functional context for why septin complex composition is tuned to the metabolic state of the cell.

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Model for the step-wise assembly of yeast septin hetero-octamers. From Weems & McMurray, eLife 2017.

  • We studied the functions of the gametogenesis-specific septin subunits in yeast and determined that they play important and unique roles in guiding the biogenesis of new membrane and cell wall. As septins also play important, but poorly understood, roles in human gametogenesis, these findings are also relevant to human septin function.

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Model for the defects in prospore membrane (PSM) biogenesis in septin-mutant yeast cells. SPB, spindle pole body; LEP, leading edge protein. From Heasley & McMurray, Molecular Biology of the Cell 2016.

  • Finally, we studied the molecular basis of action of a small molecule (forchlorfenuron, FCF) thought to act specifically on septin filament assembly in non-plant eukaryotes and therefore used by many researchers as a “septin drug”. We found clear evidence of off-target effects, using four different eukaryotic cell types.

Our studies challenge the idea that thermodynamics (i.e., the affinities between proteins randomly colliding in solution) is the main driving force of protein complex assembly, and favor largely unappreciated roles for the kinetics of protein folding and cooperativity between protein-protein interactions. We are now focusing our attention on the mechanisms by which septin proteins fold in living cells, and how the insights we gained from the study of mitotic yeast septins apply to septins in other developmental contexts, and to oligomers composed of non-septin proteins.


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Barve G, Sridhar S, Aher A, Sahani MH, Chinchwadkar S, Singh S, K N L, McMurray MA, Manjithaya R. Septins are involved at the early stages of macroautophagy in S. cerevisiae. ​Journal of Cell Science 22;131(4). pii: jcs209098. (2018) doi: 10.1242/jcs.209098.​

Weems A, McMurray M. The step-wise pathway of septin hetero-octamer assembly in budding yeast. eLife 6. pii: e23689 (2017) doi: 10.7554/eLife.23689

McMurray MA. Coupling de novo protein folding with subunit exchange into pre-formed oligomeric protein complexes: the 'heritable template' hypothesis. Biomolecular Concepts 7:271-81 (2016) doi: 10.1515/bmc-2016-0023

Heasley LR, McMurray MA. Small molecule perturbations of septins. Methods in Cell Biology 136:311-9 (2016) doi: 10.1016/bs.mcb.2016.03.013

McMurray MA. Assays for genetic dissection of septin filament assembly in yeast, from de novo folding through polymerization. Methods in Cell Biology 136:99-116 (2016) doi: 10.1016/bs.mcb.2016.03.012

Schaefer RM, Heasley LR, Odde DJ, McMurray MA. Kinetic partitioning during de novo septin filament assembly creates a critical G1 "window of opportunity" for mutant septin function. Cell Cycle 15:2441-53 (2016) doi: 10.1080/15384101.2016.1196304

Garcia G 3rd, Finnigan GC, Heasley LR, Sterling SM, Aggarwal A, Pearson CG, Nogales E, McMurray MA, Thorner J. Assembly, molecular organization, and membrane-binding properties of development-specific septins. Journal of Cell Biology 212:515-29 (2016). doi: 10.1083/jcb.201511029

Heasley LR, McMurray MA. Roles of septins in prospore membrane morphogenesis and spore wall assembly in Saccharomyces cerevisiae.Molecular Biology of the Cell 27:442-50 (2016) doi: 10.1091/mbc.E15-10-0721 

Johnson CR, Weems AD, Brewer JM, Thorner J, McMurray MA. Cytosolic chaperones mediate quality control of higher-order septin assembly in budding yeast. Molecular Biology of the Cell 26:1323-44 (2015). doi: 10.1091/mbc.E14-11-1531

Heasley LR, Garcia G 3rd, McMurray MA. Off-target effects of the septin drug forchlorfenuron on nonplant eukaryotes. Eukaryotic Cell 13:1411-20 (2014) doi: 10.1128/EC.00191-14

McMurray MA. Lean forward: Genetic analysis of temperature-sensitive mutants unfolds the secrets of oligomeric protein complex assembly. BioEssays 36:836-48 (2014)

Weems AD, Johnson CR, Argueso JL, McMurray MA. Higher-order septin assembly is driven by GTP-promoted conformational changes: evidence from unbiased mutational analysis in Saccharomyces cerevisiae. Genetics 196:711-27 (2014) doi: 10.1534/genetics.114.161182

de Val N, McMurray MA, Lam LH, Hsiung CC, Bertin A, Nogales E, Thorner J. Native cysteine residues are dispensable for the structure and function of all five yeast mitotic septins. Proteins 81:1964-79 (2013) doi: 10.1002/prot.24345

Bertin A, McMurray MA, Thorner J, Peters P, Zehr E, McDonald KL, Thai L, Pierson J, Nogales E. Three-dimensional ultrastructure of the septin filament network in Saccharomyces cerevisiae. Molecular Biology of the Cell 23:423-32 (2012) doi: 10.1091/mbc.E11-10-0850

Garcia G III, Bertin A, Li Z, Song Y, McMurray MA, Thorner J, Nogales E. Subunit-dependent modulation of septin assembly: Budding yeast septin Shs1 promotes ring and gauze formation. Journal of Cell Biology 195:993-1004 (2011)  doi: 10.1083/jcb.201107123

McMurray MA, Stefan CJ, Wemmer M, Odorizzi G, Emr SD, Thorner J. Genetic interactions with mutations affecting septin assembly reveal ESCRT functions in budding yeast cytokinesis. Biological Chemistry 392:699 (2011). doi: 10.1515/BC.2011.091

McMurray MA, Bertin A, Garcia III G, Lam L, Nogales EE and Thorner J. Septin filament formation is essential in budding yeast. Developmental Cell 20:540-9 (2011) doi: 10.1016/j.devcel.2011.02.004

Bertin A, McMurray MA, Thai L, Garcia III G, Votin V, Grob P, Allyn T, Thorner J and Nogales EE. Phosphatidylinositol-4,5-bisphosphate promotes budding yeast septin filament assembly and organization. Journal of Molecular Biology 404:711 (2010) doi: 10.1016/j.jmb.2010.10.002

Garrenton LS, Stefan C, McMurray MA, Emr SD and Thorner J. Pheromone-induced anisotropy in yeast plasma membrane phosphatidylinositol-4,5-bisphosphate distribution is required for MAPK signaling. Proceedings of the National Academy of Sciences of the United States of America 107:11805 (2010) doi: 10.1073/pnas.1005817107 

McMurray MA and Thorner J. Septins: molecular partitioning and the generation of cellular asymmetry. Cell Division 4:18. (2009) doi: 10.1186/1747-1028-4-18

McMurray MA and Thorner J. Reuse, replace, recycle: Specificity in subunit inheritance and assembly of higher-order septin structures during mitotic and meiotic division in budding yeast. Cell Cycle8(2):195-203. (2009)

McMurray MA and Thorner J. Biochemical properties and supramolecular architecture of septin hetero-oligomers and septin filaments. In: Hall PA, Russell SEG, Pringle JR, eds. The Septins. Chicester, West Sussex, UK: John Wiley & Sons, Ltd., pp. 49-100. (2008)

McMurray MA Thorner J. Septin stability and recycling during dynamic structural transitions in cell division and development. Current Biology 18:1203 (2008) doi: 10.1016/j.cub.2008.07.020

Bertin A*, McMurray MA*, Grob P*, Park SS, Garcia G 3rd, Patanwala I, Ng HL, Alber T, Thorner J, Nogales E. Saccharomyces cerevisiae septins: supramolecular organization of heterooligomers and the mechanism of filament assembly. Proceedings of the National Academy of Sciences of the United States of America 105:8274 (2008) doi: 10.1073/pnas.0803330105 *these authors contributed equally to this work.

Latest Publications in PubMed

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Current lab members

Lab Personnel
Ashley Denney, PhD student (MSTP program)
​Randi Yeager, PhD student (Molecular Biology program)
yeager.jpg


​Marc Steingesser, Professional Research Assistant (and lab dad) Steingesser.JPG

Aleyna Benson, Professional Research Assistant
Daniel Hassell, Professional Research Assistant Hassell.JPG
Ben Cooperman, PhD student (Molecular Biology program)


Former lab members

​J.P. Darling-Munson, CU Boulder undergraduate (MCDB program) JP.JPG
​Emily Singer, Professional Research Assistant Skidmore-465.JPG
Lydia Heasley, PhD (Molecular Biology Program, graduated 2016), currently postdoc in the Argueso lab in the Department of Environmental & Radiological Health Sciences at Colorado State University.
Andrew Weems, PhD (Cells, Stem Cells, and Development Program, graduated 2017), currently postdoc in the Danuser lab at the UT Southwestern Medical Center
Courtney Johnson, PRA, currently enrolled in Dental School at UC Denver
Rachel Schaefer, PRA, currently PRA in Gastroenterology, UC Denver Anschutz Medical Campus
Christina Coughlan, PhD, FCP, SI, EMT​, currently research instructor in Department of Neurology, UC Denver Anschutz Medical Campus
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The Wild Yeast Outreach Program
is funded by the National Science Foundation through award 1928900.

After >100 years of study, the budding yeast Saccharomyces cerevisiae is the best understood eukaryotic cell. A key feature that makes S. cerevisiae such a powerful tool for genetical manipulation is the efficient alternation of haploid and diploid phases (Fig. 1). Upon nutrient deprivation, most diploid S. cerevisiae strains undergo meiosis and sporulation, typically producing four haploid spores within each sporulating cell. Each spore is encased in a specialized wall that confers resistance to a variety of environmental stressors. As the two mating types reflect alternative alleles at a single locus, each meiosis produces two pairs of spores of opposite mating types, "a" and "alpha". In the lab we prevent mating between spores by physically separating them before exposing them to nutrients, which allows them to grow out from the spore wall (“germinate”) and proliferate indefinitely via budding (Fig.1). Whereas a haploid spore from most natural isolates is able to switch mating types and, via subsequent mating with one of its offspring, return to the diploid state (Fig. 1), labs use haploid strains incapable of switching. Instead, diploids are made at will by mating between haploids placed in close proximity. The ability to manipulate genes and study effects in the haploid phase and then to combine different alleles via mating/recombination/sporulation is the foundation of yeast genetics. Exploiting this life cycle in the lab context has extended our understanding of the cellular and molecular biology of S. cerevisiae to a level of detail unparalleled by any other eukaryote.
Fig1.png
Despite (or, more likely, because of) our focus on S. cerevisiae as a model for human biology, the yeast field tends to ignore the aspects of yeast biology that lack direct counterparts in human cells. Only recently have we begun to realize that in order to fully understand yeast biology, we must consider how this organism lives outside the lab.

Since in the lab germination is almost always done with isolated spores, most yeast researchers assume that if the spores are kept in contact, they will always mate. Not true! Spores frequently bud even when they are right next to a potential mating partner. Why? How does a spore decide whether to mate, or to bud?

We want to understand the circumstances in which a germinating spore buds vs mates. The research goal of this outreach program is to test the hypothesis that the tendency of a natural Saccharomyces isolate to bud vs mate upon germination can be predicted by assessing HO gene function. The HO gene is essential for mating-type switching and is only expressed in haploid cells that have budded at least once (Fig. 1). HO is thought to have evolved to allow a return to diploidy in cases where spores are dispersed. This trait is called homothallism; the failure to do so is heterothallism. If a strain never sporulates or always mates upon germination (no “lonely spores”), HO is never expressed, and mutations can, in principle, accumulate.

We hypothesize that natural Saccharomyces isolates carrying mutant HO alleles but capable of sporulation will be biased towards intra-ascus mating upon germination. To test this hypothesis, we isolate Saccharomyces species from the wild, sequence the HO locus in each, and assess sporulation ability and bud-vs-mate decision upon germination.

To engage the public in scientific research and enrich public school education in a way that also advances the research goals of our project, we will involve middle school and undergraduate students in the isolation and identification of yeast strains from the bark of oak trees or unroasted cocoa or coffee beans, and the identification of mutations in HO causing defects in mating-type switching. Here we provide a collection of resources that we are developing, which we hope will be of value to other groups undertaking similar outreach efforts.

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Description File/link​
​Intro lecture Wild Yeast Project.pdf
DNA barcoding lecture​ Species ID by DNA sequencing.pdf
​Yeast ID by microscopy quiz Wild Yeast Project: Microscopy of Bark Cultures
Map of oak yeast identified in Cheesman Park FinalCheesmanYeastMap.pdf​
 
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