Julianne H. Grose
Evaluation of academic objectives
The academic objectives of this MEG were to provide novel findings in the field of metabolic regulation while training both graduate and undergraduate students. We were able to meet these objectives as evidenced by one published scientific article, one article accepted upon minor revision, and one in preparation. In addition, fifteen oral presentations or posters were presented at scientific meetings on findings directly obtained from this MEG. Furthermore, these findings helped obtain NIH funding in 2012 that will continue through 2015, allowing for further scientific advance and training of students.
Posters/Presentations resulting from this MEG
1. DeMille, D. and Grose, JH. A Comprehensive Interactome for Yeast PAS Kinase Reveals Direct Regulation of Respiration through the Phosphorylation of Cbf1. (2013) Podium presentation. Analytical Genetics Meeting.
2. Mackay, J, DeMille, D, Grose, JH. Uncovering Regulation and Function of the Yeast NAD Kinase Utr1. (2013) Poster presentation. Analytical Genetics Meeting.
3. Badal, B. DeMille, D, Mackay, J, Grose JH. Interplay between the yeast nutrient sensing kinases Snf1, TORC1 and PAS kinase. (2013) Poster presentation. Analytical Genetics Meeting.
4. Bevard, K., Thornock, S, Collins, G., Ramsey, M. and Grose, JH. Characterizing Yeast PAS Kinase Through Random Mutagenesis (2013). Utah Undergraduate Conference for Research, Utah State University.
5. Badal, B, and Grose, JH. Snf1 Directly Phosphorylates and Activates Yeast PAS Kinase. (2013) Podium presentation, ASM Intermountain Branch Meeting, Idaho State University.
6. DeMille, D., and Grose J.H. (2012&2013) The Role of Yeast PAS kinase in PASsing Glucose. MMBIO Graduate Student Retreat, BYU.
7. DeMille, D, and Grose, JH. New Roles for PAS kinase Revealed through Protein-‐ protein Interaction Studies. (2012) Podium presentation, Intermountain Branch ASM meeting. Idaho State University. Best Biomedical Oral Presentation.
8. Findley, R, Thornock, S, Bevard, K, and Grose, JH. The Regulation of PAS kinase, a Key Sensory Kinase Required for Glucose Homeostasis (2012) Intermountain Branch ASM Meeting, Idaho State University.
9. DeMille, D, Mackay, J, Sowa, S, Hall, T, Lawrence, E, and Grose, JH. (2012) The Role of Yeast PAS kinase in Passing Glucose. Yeast Molecular Biology and Genetics Meeting, Princeton University.
10. Mackay, J, DeMille, D, Gessel A, Lawrence, E, Hall, T, and Grose, JH. (2012) A Yeast Two-‐hybrid Screen Reveals Novel Roles for Yeast PAS kinase. National Conference for Undergraduate Research (NCUR), Weber State University.
11. Findlay, R, Teng, J, Bevard, K, Thornock, S, and Grose, JH (2012) The Regulation of PAS kinase, a Key Sensory Kinase Required for Glucose Homeostasis. National Conference for Undergraduate Research (NCUR), Weber State University.
12. Grose, JH. The role of PAS kinase in PASsing cellular glucose. (2011) Analytical Genetic Meeting, Carmona, Spain.
13. DeMille, D, Mackay, J, Gessel, A, Lawrence, E, Hall, T, and Grose J.H. (2011) The Role of Yeast PAS kinase in Metabolic Regulation. Analytical Genetic Meeting, Carmona, Spain.
14. DeMille, D, and Grose JH. (2011) The Role of Yeast PAS kinase in Metabolic Regulation. (2011) ASM Intermountain Branch Meeting, Weber State University.
15. Mackay, J, Sowa, S, Loeb, S, Haines, C, and Grose JH. (2011) Finding Interacting Partners for PAS kinase. ASM Intermountain Branch Meeting, Weber State University.
Publications or articles in preparation resulting from this MEG
1. Bryan Badal, Desiree DeMille, Joseph F. Anderson, Brady Evans, Andrew Mathis, John T. Prince and Julianne H. Grose. “Interplay between the Yeast Nutrient Sensing kinases Snf1 and PAS kinase”. In preparation
2. Desiree DeMille, Jordan T. Mackay, Benjamin Bikman, Andrew D. Mathis, Steven W. Sowa, Tacie D. Hall, Benjamin Bikman, John T. Prince, and Julianne H. Grose. A Protein-‐protein Interactome for Yeast PAS Kinase 1 Reveals Direct Regulation of Respiration Through the Phosphoryltion of Cbf1. Submitted to Molecular Cell in October 2013.
3. Desiree DeMille and Julianne H Grose. PAS kinase: A Nutrient Sensing Regulator of Glucose Homeostasis. (2013) IUBMB Life 2013 Nov;65(11):921-‐9. Extramural research support obtained in 2012 from data from this MEG Molecular mechanisms of yeast PAS kinase regulation and function.
Principal Investigator: Julianne H. Grose
National Institutes of Health R15
Grant number R15 GM100376-‐01.
End date: 5/31/2015
Evaluation of the mentoring environment
We had weekly lab meetings where the students took turns presenting their findings, providing students with a broader view of science and aiding in the development of presentation skills. In addition, weekly project-specific subgroup meetings were formed where data interpretation, experimental theory and protocols, and solutions to any current experimental difficulties were discussed. Students were trained by hierarchies within these subgroups, those who have the most experience were involved in training the newer lab members. This hierarchy facilitated higher learning as students explained concepts behind their research to others and gradually acquired more independence and responsibility. All students were encouraged to take ownership of their project and to contribute by offering new avenues to explore as well as alternative experimental techniques. I was available in the lab almost every day performing experiments and have an open-door policy for the students in my lab.
Description of the results/findings of the project
PAS kinase is a recently discovered sensory kinase that is involved in physiological adaptation to nutrients. The aims of the 2011 MEG were to determine the molecular mechanisms of yeast PAS kinase regulation and function. The first aim focused on identifying and characterizing novel pathways in the regulation of PAS kinase. Our second Aim involved the characterization of putative PAS kinase substrates identified through yeast two-hybrid screens. We were successful at both of these aims as demonstrated by one published article, a article accepted with minor revisions which we hope to have revised in the next few weeks, and a third article in preparation. Each is described briefly in the corresponding Aims below.
Specific Aim 1: Identify and characterize PAS kinase regulatory mechanisms. We identified Snf1 as a putative upstream activator of PAS kinase since it was retrieved from our screen for PAS kinase substrates. We were able to demonstrate Snf1-dependent in vivo phosphorylation of yeast PAS kinase and mapped the phosphorylation sites. In addition, we provide evidence for the direct phosphorylation and activation of PAS kinase in vitro. These results are in preparation for submission to Molecular Biology of the Cell in early 2014.
Specific Aim 2: Identification and characterization of novel PAS kinase binding partners in yeast. We were able to identify 108 novel PAS kinase binding partners through large scale yeast two-hybrid and copurification screens. In addition, we characterized five of these as in vitro substrates of PAS kinase, and performed detailed in vivo characterization of one (Cbf1). These results were submitted for publication in 2013 and were accepted with minor revisions that will be submitted in January of 2014. In addition, we published a review article on the function of PAS kinase in 2013.
Description of how the 2011 budget was spent
$9,600 5 undergraduate students for 15 weeks, 8 hours per week at $8.25 per hour.This pay was for months (spring/summer) in which these students were not be enrolled in the research for credit course 494R. I chose undergraduates to be paid based on seniority in the lab and the number of hours per week they had been volunteering. The undergraduates paid in summer 2011 were Kayla Bevard, Rachel Findley, Katie Harris, Jordan Mackay, and Steve Sowa.
$9,800 Research expenses include laboratory consumables, reagents and supplies for wet lab research.
$800- DNA extraction and amplification a
$1200- 32P-ATP for kinase enzymatic assays
$2000- yeast and bacterial media, antibiotics and supplies
$2000- protein purification supplies and antibodies for detection
$1000- DNA sequencing and protein mass spectrometry
$1000- gloves, pipettes, microcentrifuge tubes, kim-wipes, parafilm, and other consumables
$600 Registration and poster fees for meeting attendance.