Signal Transduction Pathway Smorgasbord Ron Bose, MD PhD - PowerPoint PPT Presentation

Molecular Cell Biology Lecture. Oct. 24, 2017 Signal Transduction Pathway Smorgasbord Ron Bose, MD PhD Biochemistry and Molecular Cell Biology Programs Lab: 4515 McKinley Research Building, 3 rd floor Washington University School of Medicine

What is the connection? Signal Transduction

Outline 1. Nuclear Hormone Receptors 2. Cytokine Receptors – JAK/STAT Pathway 3. PI3-kinase – Akt – mTOR 4. Regulation of Protein Kinases

Resources: Nuclear Hormone Receptors https://www.nursa.org/nursa/index.jsf Online Course: https://www.nursa.org/nursa/flashTutorial/gene/nu clearReceptor/start.jsf Reference: McKenna NJ and O'Malley BW. An interactive course in nuclear receptor signaling: concepts and models. Sci STKE. 2005 , tr22.

Nuclear Hormone Receptor Superfamily 1. 48 Human genes 2. Major Categories: Thyroid Hormone TR, RAR, PPAR, Vitamin D receptor, LiverX Receptor (TR)- like Receptor Estrogen Receptor (ER)- ER, PR, AR, Estrogen Receptor Related, like Glucocorticoid receptor, Mineralocorticoid receptor Retinoid X Receptor (RXR) RXR, Hepatocyte nuclear factor-4, etc. like Knock-out in mice causes reproductive, developmental, or metabolic abnormalities.

AF: Activation Function. Mediate transcriptional activation DNA Binding Domain Ligand Binding Domain www.nursa.org/

AF: Activation Function. Mediate transcriptional activation www.nursa.org/

Bind as homodimers Bind as heterodimers with RXR Hormone response elements Hormone response elements are inverted repeats. are direct repeats. www.nursa.org/

Ligand Present Ligand Absent www.nursa.org/

Movie: https://nursa.org/nursa/about/tutorial.jsf Tab 12. Nuclear Hormone Action Model www.nursa.org/

Outline 1. Nuclear Hormone Receptors 2. Cytokine Receptors – JAK/STAT Pathway 3. PI3-kinase – Akt – mTOR 4. Regulation of Protein Kinases

Cytokine Receptors – JAK/STAT Pathway Baker et al., Oncogene (2007) 26, 6724–6737

Cytokine Receptors Baker et al., Oncogene (2007) 26, 6724–6737

JAK = Janus kinases 4 genes in humans and mice • TYK2 (first gene in this family to be identified) • JAK1, JAK2, JAK3 Baker et al., Oncogene (2007) 26, 6724–6737

STAT= Signal Transducers and Activators of Transcription Baker et al., Oncogene (2007) 26, 6724–6737

Cytokine Receptors – JAK/STAT Pathway Baker et al., Oncogene (2007) 26, 6724–6737

Recptor Tyrosine Kinases Examples– EGFR, Her2, etc from Marmor, Skaria, and Yarden 2004

Outline 1. Nuclear Hormone Receptors 2. Cytokine Receptors – JAK/STAT Pathway 3. PI3-kinase – Akt – mTOR 4. Regulation of Protein Kinases

PI3-kinase – Akt – mTOR Songet al., Nature Rev Mol Cell Bio 2012

PI3-kinase – Akt PI3K PtdIns(4,5)P 2 PtdIns(3,4,5)P 3 (PIP2) (PIP3) PTEN PDK1 Akt Zoncu et al., Nature Rev Mol Cell Bio 2011

mTOR complexes mTORC1 Rapamycin sensitive Responds to nutrient level, growth factors, energy, and stress. mTORC2 NOT rapamycin sensitive Inputs into mTORC2 less well known. Zoncu et al., Nature Rev Mol Cell Bio 2011

mTORC1 substrates • S6 kinase 1 (S6K1) • eIF-4E binding protein (4E-BP) Zoncu et al., Nature Rev Mol Cell Bio 2011

mTORC2 substrates Akt mTORC2 PDK1 1 2 T308 S473 PH Kinase Domain domain 3 Downstream substrates: TSC complex, PRAS40, etc. Zoncu et al., Nature Rev Mol Cell Bio 2011

Bringing it all together mTOR is a signal integrator, like the chips and circuits in your smart phone Zoncu et al., Nature Rev Mol Cell Bio 2011

Outline 1. Nuclear Hormone Receptors 2. Cytokine Receptors – JAK/STAT Pathway 3. PI3-kinase – Akt – mTOR 4. Regulation of Protein Kinases

More information available at: http://kinase.com/web/current/ Manning et al., Science 2002

Regulation of Protein Kinases 1. Post-translation modifications.  Phosphorylation-dependent  Activation Loop 2. Protein-protein interactions  Regulatory Subunits  Dimers

Structure of PKA catalytic domain α Helices β Sheets N-lobe C Helix C-lobe Caplan, Science STKE 2005

Structural features of the PKA Activation Loop Illustration from Nolen et al, Mol. Cell, Vol. 15, p.661-675, 2004

Phosphorylation of the MAP Kinase activation loop • Phosphorylation on threonine and tyrosine • Phospho-Thr 183 contacts α -C and promotes active conformation • Phospho-Thr 183 promotes ERK2 dimerization via conformational changes in C-terminal extension AKT phosphorylation at Illustration taken from Huse and Kuriyan, Cell 109, 275-282 (2002) T308 is also Activation Loop Phosphorylation

MAP Kinase Structure Thr183 Tyr185 Unphosphorylated Phosphorylated Canagarajah et al Cell 90, 859-869 (1997)

MAP Kinase Structure Unphosphorylated Thr183 Phosphorylated Tyr185 Canagarajah et al Cell 90, 859-869 (1997)

Cyclin - Cyclin-dependent kinase (CDK) Complex Cyclin A Cyclin-dependent kinase (Cdk2) Jeffrey et al Nature 376, 313-320 (1995)

Jeffrey et al Nature 376, 313-320 (1995) C-helix C-helix Activation Loop Activation Loop Cdk2 Cdk2.CyclinA

Asymmetric Dimer Formed by the EGFR Kinase Domain EGFR kinase domain asymmetric dimer Zhang, Gureasko, Shen, Cole, and Kuriyan. Cell 2006

Summary 1. Nuclear hormone receptors consist of a DNA- binding domain and ligand-binding domain. 2. Cytokine receptors signal through the JAK kinases, which have 2 kinase domains, and the STAT transcription factors. 3. mTOR is a signal integrator for metabolic and growth factor signaling. 4. Protein kinases are regulated by PTM’s and protein-protein interactions.