From Membrane to Nucleus: Teaching Cell Signaling

via the Ras Pathway

CELL BIOLOGY TEAM

Kathleen Marrs, IUPUIJames Marrs, IUPUIMariah Judd, IUPUITodd Murphy, Ivy Tech Community College (IUPUI)Robert Steven, University of ToledoTomer Avidor-Reiss, University of ToledoHenry Chang, Purdue University

Facilitators:Brian Kinkle, University of Cincinnati Sadie Hebert, University of Minnesota

CONTEXT

0This unit was prepared for a sophomore level cell biology course, taught mid way through the semester0 This unit was designed with the flexibility to be scaled

down to a freshman level biology course.0Unit designed to take 1-2 (one hour) class sessions

EXPECTED BACKGROUND

0Basics of cell structure and organelles 0Protein control by phosphorylation 0Understand that cells have the ability to respond to

extracellular stimuli0Basics of cell signaling cascades (including various

types of G-proteins)

TEACHABLE UNIT GOALS AND OUTCOMES

I. Know specific components and relationship between components of the Ras pathway affecting cell proliferation.

a) Order the sequence of proteins involved in the Ras pathway and describe their cellular location (BT 1-2)

b) Describe the role of each component of the Ras pathway (BT 2-3)

II. Evaluate the consequences of perturbations to the signaling pathway using real data.

a) Analyze the effect of mutations on the components of the Ras pathway (BT 5-6)

b) Practice the skill of analyzing data, controls and outcomes of experiments. (BT 4-5)

TEACHABLE UNIT GOALS AND OUTCOMES

I. Know specific components and relationship between components of the Ras pathway affecting cell proliferation.

a) Create a schematic depicting the RTK/RAS/MAP kinase signaling pathway in both the active and inactive states. (BT 1-2)

b) Discuss the role and significance of each component of the Ras pathway. (BT 2-3)

II. Evaluate the consequences of perturbations to the signaling pathway using real data.

a) Analyze the effect of perturbation(s) on the components of the Ras pathway. (BT 4-5)

b) Develop the skill of analyzing data, controls and outcomes of experiments. (BT 4-5)

(BT=Bloom’s Taxonomy)

PRE-ASSIGNMENTS

0Read the introduction from “The History of Ras: the Saga Continues” by Cox and Der, 2010

0Review chapter section in text book that covers Ras pathway and review section questions

0MAPK pathway video by Genentech (Youtube video) with video reflections

PRE-ASSIGNMENTS

0Read the introduction from “The History of Ras: the Saga Continues” by Cox and Der, 2010

0Review chapter section in text book that covers Ras pathway and review section questions

0MAPK pathway video by Genentech (Youtube video) with video reflections

PRE-ASSIGNMENTS SUMMARY

Modulation of signalling by Sprouty: a developing storyHong Joo Kim & Dafna Bar-SagiNature Reviews Molecular Cell Biology 5, 441-450 (June 2004) doi:10.1038/nrm1400

From Membrane to Nucleus:

Teaching Cell Signaling via the Ras Pathway

Our Tidbit

Activation of a G-protein involves ___________.

a) Cleavage of the G-proteinb) Methylation of the G-proteinc) Binding of GTPd) Binding of GDPe) Techno music

Clicker Question

Ras is a __________.

a) Signal transduction moleculeb) Transcription factorc) Calcium channeld) Membrane receptore) Flavor of the Month at Starbucks

Clicker Question

WHY DO WE CARE ABOUT RAS?

0Ras is the most common oncogene in human cancer.0 90% of all pancreatic cancers0 25% of all human tumors

0The Ras pathway provides a focus on the origins of cancer at the molecular level.

0Ras is a key cellular protein for cell growth, differentiation and survival.

http://www.sciencephoto.com/

RAS-ZLE DAZZLE PUZZLE

Materials needed:0 One 8.5x11 puzzle board0 2 Envelopes labeled #1 “OFF” and #2 “ON” filled with puzzle

pieces

Working with your pod…0 First: read and open envelope #1 “OFF” and assemble the pieces

in the appropriate place on the puzzle board. (5 minutes)0 Second: share your puzzle with the other pods at your table to

compare and discuss. (5 minutes)

Ras pathway modeling activity

RAS-ZLE DAZZLE PUZZLE

“OFF” State

RAS-ZLE DAZZLE PUZZLE

Working with your pod…0 Third: read and open envelope #2 “ON” and add the new pieces

to the puzzle board. (5 minutes) 0 Fourth: share your completed puzzle with the other pods at your

table to compare and discuss. (5 minutes)

Ras pathway modeling activity

RAS-ZLE DAZZLE PUZZLE

“ON” State

What initially triggers a change in gene expression (in a WT cell)?

a) Ras Activationb) GTP Hydrolysisc) MAP Kinase Phosphorylationd) Growth Factor – Receptor binding

Clicker Question

RAS-ZLE DAZZLE PUZZLE:WHAT IS THE SIGNIFICANCE?

0What is the significance and outcome of…

0 the receptor – ligand interaction0 the Ras activation0 the MAP kinase cascade

0What is the significance of the ERK translocation and what is one possible outcome?

WHEN GOOD CELLS GO BAD:DISRUPTIONS IN THE PATHWAY

0What happens when we perturb the pathway?

Summative Assessment

SUMMATIVE HOMEWORK

0Practice with data analysis, controls and outcomes.

0Uses real data from primary literature.

Summative Assessment

RITA LEVI-MONTALCINI

“Above all, don’t fear difficult moments. The best comes from them.”

In Praise of Imperfection, Levi-Montalcini’s biography

0Nobel laureate0Discovered growth

factor signaling0 Italian Jewish

scientist0Lived to age 103

Image

TIDBITGOALS AND OUTCOMES

Know specific components and relationship between components of the Ras pathway affecting cell

Create a schematic depicting the RTK/RAS/MAP kinase signaling pathway in both the active and inactive states.(BT 1-2)

Discuss the role and significance of each component of the Ras pathway(BT 2-3)

Goal I

Outcome a)

Outcome b)

With thanks to HHMI, The University of Minnesotaand the National Academies NorthStar (Midwest) Summer Institute (NANSI):

Thanks also to

Brian and Sadie,

our amazing

facilitators