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PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle ([email protected] )

PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle ([email protected])[email protected]

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Page 1: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

PLS 623: Physiology of Plants II

Spring Semester 2014

Jan Smalle ([email protected])

Page 2: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Journal club format

Why?

- Practice how to prepare papers and express ideas correctly.

- Training in science debate.

- Practice presentation: research seminar.

Page 3: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu
Page 4: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu
Page 5: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

TARGETED PROTEIN DEGRADATION

Lectures: January 15, 17

Exam: January 22

Page 6: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu
Page 7: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu
Page 8: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Central dogma of genetics

Page 9: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Central dogma of genetics

Regulation of an organisms differentiation and responses to the environment are the result of changes in gene expression.

OLD THEORY: Regulation of gene expression is predominantly controlled at the transcription level.

NEW THEORY: Regulation of gene expression is the result of a combination of protein synthesis (transcription/translation) and protein degradation controls.

Page 10: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

DNA RNA PROTEIN

Light

Light responses

EXAMPLE Day/night cycle: day light activates gene expression

Page 11: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

DNA RNA PROTEIN ????

Dark

Light responses ?

Day/night cycle: During night time, light-inducible genes are down-regulated

However, regulatory proteins that promote the light response need to be removed in the dark. Otherwise, they would interfere with the dark response.

Page 12: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

PhotomorphogenesisHY5: a transcriptional activator of photomorphogenic development

5 days old Arabidopsis seedlings grown under a range of light intensities

Light intensity

(Osterlund et al., 2000)

Western blot using HY5 antiserum

Page 13: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Ethylene signalingEIN3: a transcriptional activator of ethylene responses

4 days old Arabidopsis seedlings grown in the dark on a range of concentrations of the ethylene precursor ACC

(Guo and Ecker, 2003)

Western blot using EIN3 antiserum

Page 14: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Proteolysis

Errors: Misfolded: Cleaved: AA supply: Regulation:

PRQTEIN PRO

T

EIN

PRO + PROTEIN

PROTEIN

PR

OT

E

IN

PROPROTEIN

PR

QT

E

IN P

R

OT

E

IN

PROTEIN

PROTEIN

PR

OT

E

IN

Page 15: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Non-specific Proteolysis

PROTEIN

PR

OT

E

IN

PROTEIN

PR

OT

E

IN P

R

OT

E

IN

PROTEIN PROTEIN

PR

OT

E

IN

PROTEIN

PR

OT

E

IN

PROTEIN

PR

OT

E

IN

Page 16: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Specific or Targeted Proteolysis

PROTEIN PROTEIN PROTEIN

PR

OT

E

IN

PROTEIN PROTEINPROTEIN

Page 17: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Conditional Proteolysis

PROTEIN PROTEIN PROTEIN

PR

OT

E

IN

PROTEIN PROTEINPROTEIN

DARK

Page 18: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Conditional Proteolysis

PROTEIN PROTEIN*Kinase

Phosphatase

PRO

TE

IN

Many cases of conditional proteolysis involve secondary modifications of target proteins. For example: phosphorylation/dephosphorylation.

Page 19: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Conditional Proteolysis

PROTEIN PROTEIN*Kinase

Phosphatase

PRO

TE

IN

Many cases of conditional proteolysis involve secondary modifications of target proteins. For example: phosphorylation/dephosphorylation.

Page 20: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

Conditional Proteolysis

PROTEIN PROTEIN PROTEIN

PR

OT

E

IN

PROTEIN PROTEINPROTEIN

Signal

Receptor

*

X?

Page 21: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu
Page 22: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu
Page 23: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu
Page 24: PLS 623: Physiology of Plants II Spring Semester 2014 Jan Smalle (jsmalle@uky.edu)jsmalle@uky.edu

[Vierstra, 2003 TIPS]