switch from systems microbiology to systems cell biology...

8
1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election day) 2 L14: 14 Lectures past and 11 to go Part I: ‘The cell as a well-stirred bioreactor’ topics - Lambda phage lysis-lysogeny switch - Synthetic genetic switch - Switches as memory storage - Chemotaxis: perfect adaptation or not? - Synthetic genetic oscillators main assumption - well-stirred reactor - absence of chemical gradients (justification: small cells, diffusion mixes) 3 Switch from Systems Microbiology to Systems Cell Biology The importance of diffusion and gradients for cellular regulation 4 Part II: Systems Cell Biology ‘Importance of gradients and diffusion in cellular regulation’ After two lectures on diffusion and reaction-diffusion equations, we will focus during the coming lectures on the following biological systems: 1. Eukaryotic chemotaxis Gradient sensing in Dictyostelium (aka‘amoeba’) 2. Computing the middle of a cell Pole-to-pole oscillations in Escherichia coli 3. Self organization of the cytoskeleton

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Page 1: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

1

Due

dat

e fo

r PS

#3 h

as b

een

exte

nded

to

Thur

sday

11/

04/0

4(a

void

ing

elec

tion

day)

2

L14:

14

Lect

ures

pas

t and

11

to g

o

Part

I: ‘T

he c

ell a

s a

wel

l-stir

red

bior

eact

or’

topi

cs

- Lam

bda

phag

e ly

sis-

lyso

geny

sw

itch

- Syn

thet

ic g

enet

ic s

witc

h- S

witc

hes

as m

emor

y st

orag

e- C

hem

otax

is: p

erfe

ct a

dapt

atio

n or

not

?- S

ynth

etic

gen

etic

osc

illat

ors

mai

n as

sum

ptio

n

- wel

l-stir

red

reac

tor

- abs

ence

of c

hem

ical

gra

dien

ts(ju

stifi

catio

n: s

mal

l cel

ls, d

iffus

ion

mix

es)

3

Sw

itch

from

Sys

tem

s M

icro

biol

ogy

toS

yste

ms

Cel

l Bio

logy

The

impo

rtanc

e of

diff

usio

n an

d gr

adie

nts

for c

ellu

lar r

egul

atio

n

4

Part

II: S

yste

ms

Cel

l Bio

logy

‘Impo

rtanc

e of

gra

dien

ts a

nd d

iffus

ion

in c

ellu

lar r

egul

atio

n’A

fter t

wo

lect

ures

on

diffu

sion

and

reac

tion-

diffu

sion

equ

atio

ns,

we

will

focu

s du

ring

the

com

ing

lect

ures

on

the

follo

win

gbi

olog

ical

sys

tem

s:

1. E

ukar

yotic

che

mot

axis

Gra

dien

t sen

sing

in D

icty

oste

lium

(aka

‘am

oeba

’)2.

Com

putin

g th

e m

iddl

e of

a c

ell

Pol

e-to

-pol

e os

cilla

tions

in E

sche

richi

a co

li

3. S

elf o

rgan

izat

ion

of th

e cy

tosk

elet

on

Page 2: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

5

Prok

aryo

tic v

ersu

s eu

kary

otic

cel

ls

e.g

E. c

oli,

cyan

obac

teria

~ 10

6 bp,

~ 5

000

gene

s6

Prok

aryo

tic v

ersu

s eu

kary

otic

cel

ls

e.g.

yea

st, a

moe

ba,

plan

ts, f

lies,

m

an &

mic

e ...

~ 10

9 bp,

~ 4

0000

gen

es !!

7

Topi

c I:

Spat

ial o

scill

atio

n in

E. c

oli

sim

ilar t

o ge

netic

osc

illat

ors,

but

now

we

cann

ot ig

nore

the

spat

ial d

imen

sion

s

biol

ogic

al fu

nctio

n:

dete

rmin

e th

e ce

nter

of t

he c

ell,

to p

repa

re fo

r pro

per c

ell d

ivis

ion

8

Page 3: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

9gf

p-m

inC

10

GFP-minD

11gf

p-m

inE

is lo

caliz

edin

a ri

ng

minE

minD

12

gfp-minE

Page 4: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

1314

Rec

ent r

esul

ts d

emon

stra

teth

at th

e m

in p

rote

ins

asse

mbl

e in

helic

es

15

Topi

c II:

Gra

dien

t sen

sing

my

favo

rite

mov

ie ..

.

neut

roph

il ch

asin

ga

bact

eriu

m

Dic

tyos

teliu

m (a

moe

ba)

sens

ing

a gr

adie

nt o

fcA

MP

sim

ilar t

o ch

emot

axis

of E

. col

i, bu

t now

we

cann

ot ig

nore

the

spat

ial d

imen

sion

s

16

Page 5: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

17

cycl

ic A

MP

(cA

MP

) is

an a

ttrac

tant

for D

icty

oste

lium

(soc

ial a

moe

ba)

18

uni

form

ste

p in

cA

MP

cA

MP

gra

dien

t

19

Res

pons

e of

Dic

tyos

teliu

m to

cA

MP

initi

aldi

strib

utio

nt ~

3 s

stea

dy-s

tate

dist

ribut

ion

t ∞

unifo

rm s

tep

in c

AM

P

unifo

rm a

nd tr

ansi

ent

pola

rized

and

per

sist

ent

cAM

P g

radi

ent

20

Res

pons

e of

Dic

tyos

teliu

m to

pul

sed

cAM

P

Page 6: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

21

geom

etry

of c

ell:

circ

ular

insi

de c

ytop

lasm

: wel

l-stir

red

insi

de m

embr

ane:

diff

usio

n-lim

ited

GFP

-PH

bin

ds s

peci

al li

pids

in m

embr

ane:

PIP

2 an

d P

IP3

22

The

mol

ecul

es in

the

mod

el:

23

aggr

egat

ion

of a

pop

ulat

ion

of a

moe

ba

biol

ogic

al fu

nctio

n:de

velo

pmen

t of a

mul

ticel

lula

r stru

ctur

e24

PH-GFP

powerful reporter

Page 7: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

25

Topi

c III

: Sel

f-org

aniz

atio

n of

the

cyto

skel

eton

Cen

ter f

indi

ng in

an

euka

ryot

ic c

ell:

fissi

on y

east

The

impo

rtanc

e of

the

cyto

skel

eton

26

27C

hang

et a

l.28

Pos

itive

feed

back

as

a m

echa

nism

sfo

r spo

ntan

eous

cel

l pol

ariz

atio

n ?

Page 8: Switch from Systems Microbiology to Systems Cell Biology ...web.mit.edu/biophysics/sbio/PDFs/L14_notes.pdf1 Due date for PS#3 has been extended to Thursday 11/04/04 (avoiding election

29

Cel

ls a

rres

ted

in G

1

Ove

rexp

ress

ed c

dc42

-GFP

(dom

inan

tly a

ctiv

e)

All

inte

rnal

and

ext

erna

lcu

es re

mov

ed

1 ho

ur a

fter i

nduc

tion

of c

dc42

-gfp

3 ho

ur a

fter i

nduc

tion

of c

dc42

-gfp

Wed

lich-

Sol

dner

et a

l.S

cien

ce 2

99, 1

231

(200

3)30

dyna

mic

s of

sym

met

rybr

eaki

ng

31

Pos

itive

feed

back

thro

ugh

cyto

skel

etal

med

iate

d cd

c42

trans

port

32

mor

e in

itial

cdc

42 o

n m

embr

ane,

mor

e ca

psst

rong

er fe

edba

ck, l

ess

caps

mod

el:

expe

rimen

ts: