Robert Youker

Department of Biology

Assistant Professor of Molecular Biology

Office: Natural Science Building 128
Phone: 828-227-2945
Email: rtyouker@wcu.edu

Education and Training:

Postdoctoral Training at University of Pittsburgh School of Medicine

Postdoctoral Training at Vollum Institute

Ph.D. in Molecular, Cellular & Developmental Biology (Teaching minor), University of Pittsburgh, 2005

M.H.S in Reproductive Biology, Johns Hopkins University, 2001

B.S. in Biochemistry, Manhattan College, 1998

Courses Taught:

BIOL 333: Cell and Molecular Biology

Research Description/Interests:

In the 1960s, Gregorio Weber observed fluctuations in the native structures of proteins using innovative fluorescence spectroscopic techniques. He described proteins in solution as "kicking and screaming stochastic molecules" and subsequent biophysical studies have upheld his view. Today, Proteins are known to be dynamic molecules that can change shape on broad timescales (nanoseconds to seconds) and these changes can encompass angstrom to nanometer distances. These changes in shape lead to rearrangements in protein-protein interactions and composition of protein complexes. Protein dynamics are necessary for a variety of cellular processes that include protein trafficking, signal transduction and protein folding.

My laboratory is interested in how protein dynamics, especially the process of oligomerization, regulates the sorting of membrane proteins in polarized epithelial cells under normal and disease conditions. We are also interested in how oligomeriztion affects G-protein coupled receptor (GPCR) signaling. Finally, we are studying the effects of inappropriate protein dynamics that occur during protein misfolding and can lead to human disease such as Cystic Fibrosis.

We use a combination of molecular, biochemical, and advanced microscopy imaging techniques to study these diverse cellular events. Students in my laboratory will learn mammalian/yeast cell culture, molecular cloning techniques, basic/advanced fluorescence microscopy, and data analysis of microscopic images.

Publications:

Youker RT* and Teng H. (2014) Measuring Protein Dynamics in Live Cells: Protocols and Practical Considerations for Fluorescence Fluctuation Microscopy. (accepted) Journal of Biomedical Optics. * Corresponding author

Labilloy A, Youker RT, Bruns JR, Kukic I , Kiselyov K , Halfter W, Finegold D, Hadad do Monte SJ, Weisz OA. (2014) Altered Dynamics of a Lipid Raft Associated Protein in a Kidney Model of Fabry Disease. Mol. Gen. Metab. 111: 184-192. PMCID: PMC3946758

Youker RT *, Bruns JR, Costa SA, Rbaibi Y, Lanni F, Kashlan OB, Teng H, and Weisz OA. (2013) Multiple motifs regulate apical sorting of p75 via a mechanism that involves dimerization and higher-order oligomerization. Mol. Biol. Cell. 24: 1996-2007. PMCID: PMC3681702. * Corresponding author

Mo D, Costa SA, Ihrke G, Youker RT, Pastor-Soler N, Hughey RP, and Weisz OA. (2012) Sialylation of N-linked glycans mediates apical delivery of endolyn in MDCK cells via a galectin-9 dependent mechanism. Mol. Biol. Cell. 23: 3636-46. PMCID: PMC3442411

Hutt DM, Martino Roth D, Chalfant M, Youker RT, Matteson J, Brodsky JL, Balch WE (2012) FKBP8 Peptidyl-Prolyl-isomerase activity manages a late stage of CFTR folding and stability. J. Biol. Chem. 287: 21914-25. PMCID: PMC3381152

Mattila PE, Youker RT, Mo D, Bruns JR, Cresawn KO, Hughey RP, Ihrke G, and Weisz OA. (2012) Multiple biosynthetic trafficking routes for apically secreted proteins in MDCK cells. Traffic. 13: 433-442. PMCID: PMC3276681

Youker RT, Shinde U, Day R and Thomas G (2009) At the Crossroads of Homeostasis and Disease: Roles of the PACS proteins in Membrane Traffic and Apoptosis. Biochem. J. 421: 1-15. PMID: 19505291

Aslan JE, You HH, Williamson DM, Endig J, Youker RT, Thomas L, Shu HJ, Du YH, Milewski RL, Brush MH, Possemato A, Sprott K, Fu H, Greis KD, Runckel DN, Vogel A, Thomas G (2009) Akt and 14-3-3 Control a PACS-2 Homeostatic Switch that Integrates Membrane Traffic with TRAIL-Induced Apoptosis. Mol. Cell 34: 497-509. PMCID: PMC2744858

Atkins KM, Thomas L, Youker RT, Harriff MJ, Pissani F, You H, Thomas G (2008) HIV-1 Nef binds PACS-2 to Assemble a Multikinase Cascade that Triggers Major Histocompatibility Complex Class I (MHC-I) Down-regulation: Analysis using short interfering RNA and knock-out mice. J. Biol. Chem. 283: 11772-84. PMCID: PMC2431057

Youker RT and Brodsky JL (2007) Regulation of Hsp70 Function: Hsp40 Co-Chaperones and Nucleotide Exchange Factors in Cell Stress Proteins, Calderwood SK (ed), Springer Science Publishers, New York. 209-227

Zhang H, Schmidt BZ, Sun F, Condliffe SB, Butterworth MB, Youker RT, Brodsky JL, Aridor M, Frizzell RA (2006) Cysteine String Protein monitors late steps in Cystic Fibrosis Transmembrane Conductance Regulator Biogenesis. J. Biol. Chem. 281: 11312-11321. PMID: 16469739

Youker RT, Walsh P, Beilharz T, Lithgow T and Brodsky JL (2004) Distinct Roles for the Hsp40 and Hsp90 Molecular Chaperones during Cystic Fibrosis Transmembrane Conductance Regulator Degradation in Yeast. Mol. Biol. Cell 15: 4787-4797. PMCID: PMC524727

Sullivan ML, Youker RT, Watkins SC, Brodsky JL (2003) Localization of the BiP molecular chaperone with respect to endoplasmic reticulum foci containing the cystic fibrosis transmembrane conductance regulator in yeast. J. Histochem. Chytochem. 51: 545-548. PMID: 12642634

Akingbemi BT, Youker RT, Sottas CM, Ge R, Katz E, Klinefelter GR, Zirkin BR, Hardy MP (2001) Modulation of the rat Leydig cell steroidogenic function by di (2-ethylhexyl) phthalate. Biol. Reprod. 65: 1252-1259. PMID: 11566751

Trentacoste SV, Friedmann AS, Youker RT, Breckenridge CB, Zirkin BR (2001) Atrazine effects on testosterone levels and androgen-dependent reproductive organs in peripubertal male rats. J. Androl. 22:142-148. PMID: 11191080

Invited Presentations:

9th International Weber Symposium on Innovative Fluorescence Methodologies in Biochemistry and Medicine, Lihue, Kauai, HI, June 15-20, 2014
Short Talk: Proteasome inhibition alters dynamics of CFTR aggregation measured by N&B analysis

11th Annual Pittsburgh Symposium on Membrane Traffic, Pittsburgh, PA, May 3, 2012
Fellow Presentation: Multiple motifs direct the apical sorting of a model raft-independent protein

Seminar at Nankai University, Tianjin, China, March 2011
Graduate student lecture: Animal models of kidney disease

Seminar at Slippery Rock University, Slippery Rock, PA, October 15, 2010
Biology Seminar Series: Maintaining cell polarity - Which way to the apical membrane?

Poster Presentations (Partial list):

9th International Weber Symposium on Innovative Fluorescence Methodologies in Biochemistry and Medicine, Lihue, Kauai, HI, June 15-20, 2014. Biophysical studies to dissect the role of oligomerization in apical protein sorting. Youker RT, Bruns JR, Weisz OA.

Gordon Research Conference in Lysosomal Diseases, Lucca, Italy, April 14-19, 2013. Dynamics of a raft associated protein in a live cell model of Fabry disease. Labilloy A, Youker RT, Bruns JR, Kukic I, Kiselyov K, Halfter W, Finegold D, Weisz OA.

ASBMB special symposia, Biochemistry of Membrane Traffic, Tahoe City, CA, October 28-31, 2010. Dissecting the roles of O-glycosylation and dimerization in the apical sorting of a model raft-independent protein . Youker RT, Lanni F, Teng H and Weisz OA.

44th Annual American Society of Cell Biology Meeting, Washington DC, December 3-8, 2004. The Hsp40 and Hsp90 molecular chaperones exhibit opposing effects on the ER Associated Degradation (ERAD) of CFTR in S. cerevisiae. Youker RT, Walsh P, Beilharz T, Ryan K, Lithgow T, and Brodsky JL.. Abstract 2451, published as Supplement to Mol. Biol. Cell, 15 (12) 2004.

HUPO 2nd Annual & IUBMB XIX World Congress, Montreal, Canada, October 8-11, 2003. The Role of Hsp90/Hsp90 complex in the degradation of CFTR in S. cerevisiae. Youker RT and Brodsky JL. Abstract 42.14, published in Mol. & Cellular Proteomics, 2 (9) 2003.

16th Annual North American Cystic Fibrosis Conference, New Orleans, Louisiana, October 3-6, 2002. Hsp70 and Hsp90 chaperones have opposing roles in the Degradation of CFTR in S. cerevisiae. Youker RT, Scott CM, and Brodsky JL. Abstract 40, published in Pediatric Pulmonology, S24 (34) 2002.







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