Are cyclic AMP effects related to real physiological phenomena?

Vol. 57, No. 2, 1974

ARE

BIOCHEMICAL

CYCLIC

AMP

EFFECTS

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

RELATED

TO REAL

Agnes

January

25,

-

that cyclic

its metabolism are absent from

Bacillus

exhibits

cyclic

is

mediated

catabolite

currently by

AMP.

AMP and enzymes involved

in

Cl), this organism

megaterium

repression which can be ZargeZy relieved

it

level shown

amounts

seemed

of

Materials

of

and

source.

by

The bacteria IO-‘N

with

50 pg/ml

tosidase

that

specifically

also

as

binds

dependent

BaciZZus megateriwn Adenyl

(
to be absent

to investigate

in Bacilk

is cyclic

on

regu-

a

AMP.

AMP

appear

bacteria

in

does

cyclaee

in this

the effect

not and

organism.

of cyclic

AMP

megateriwn.

Methods.

by Aubert

with

of cyclic

cyclic

BaciZZus megaterium strain described

that

repression

111

interest

repression

catabolite

factor

catabolite

recently

repression

catabolite

a protein

phosphodiesterase

AMP

Therefore

and

interpret

detectable

cyclic

that

intracellular

has been

contain

AMP

authors

of the It

believed

cyclic

Most

lation

as

Cellulaire PARIS

AMP.

It

on

?

19'74

SUMRY - In spite of the fact

still

PHENOMENA

ULLMANN

Service de Biochimie INSTITUT PASTEUR

Received

PHYSIOLOGICAL

galactose.

and Millet were After

chloramphenicol

was assayed

MA102

grown 3-4

grown

(21 in the at 30°C.

in

cell

(31.

salt

of 0.2

B-galactosidase

suspension

previously

minimal

presence

generations,

and the

as described

was

growth

medium

% of a carbon

was

induced

was arrested

was sonicated.

@-galac-

Vol. 57, No. 2, 1974

BIOCHEMICAL

AND BIOPHYSICAL RESEARCfl COMMUNICATIONS

Results.

Table glucose,

I

shows

the

glycerol

and

differential

rate

galactose

as

of

g-galactosidase

sources

carbon

TABLE

DIFFERENTIAL

RATE

OF

synthesis

and

the

effect

of

I

@-GALACTOSIDASE

SYNTHESIS

U/mg

source

Addition

9.3 +cAMP

8.6

79

Glycerol +cAMP

280

+S'AMP

210 +cAMP

820

+S'AMP

220

3.9

I.

were

Cultures Generation

time :

9

3.6

73

Galactose

of

2.9

27

+5'AMP

tose

+cAmP - cfiw

Ratio

B-galactosidase

Glucose

TABLE

cyclic

BACILLUSMEGATERIUM

IN

Carbon

in

135

min.

in

grown

and

glucose Cyclic

: AMP

induced

as

85

;

and

min. S'AMP

described in

were

mm.

349

glycerol used

Muteriak

in : at

100

a final

min.

and Methods. ;

in

concentration

galac-

BIOCHEMICAL

Vol. 57, No. 2, 1974

AMP and S'AMP on the dase synthesis times

is

higher

concluded

in

that

pression.

rate

about galactose

Cyclic

times

than

it

synthesis

seems to be specific

is

1 shows the

can be seen that

higher

in glycerol

in glucose. under

the

of the

nature

cyclic

AMP.

for

It

is

AMP stimulates

of 3 to 4 independently

Figure

of synthesis. eight

enzyme

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

the

rate

carbon

I IO

twenty can be

of catabolite

by a factor

source.

of the

re-

The effect

cyclic

AMP effect.

I c 20 CAMP

1.

Differential cyclic time

it

of enzyme synthesis

mM

Figure

Therefore

dependence

I

and about

control

of the

concentration

g-galactosi-

rate

of f3-galactosidase

AMP concentration. is

80 minutes

Maximal

effect

fairly

high

giving

maximal

The carbon

[independent

is

obtained

although,

in

effects

even at 17 mM cyclic

synthesis source

is

glycerol

of the presence

at about fact,

it

in Eschetichia AMP bacterial

only

about

coli. growth

It is

not

AMP which twice

of

and the

or absence

IO mM cyclic is

as a function

generation

of cyclic

AMPI.

may appear

the concentration

has to be noted

that

affected.

Discussion. Even though of cyclic

it

appears

AMP in relieving

quantitatively catabolite

quite repression

350

significant, in BaeiZZus

the

effect

megaterium

Vol. 57, No. 2, 1974

BIOCHEMICAL

cannot

be

cyclic

AMP

as

well

as

cyclic

AMP

in

fact

interferes

must

considered

"physiological",

A possible

fered nucleotide

cyclic

in

BaciZZus

However

gated.

presence

in

AMP,

the

if

cyclic

whose

level

GMP

or

lack

conclude

normal

effects

GNP. not

AMP

its

the

to

that

control,

which

AMP.

has

for

appears

therefore

with

cyclic

megaterilnm

GWP,

way

a mediator be

organism

should

cyclic

could

test

of cyclic

some

than

for

by

this

We

other

candidate

with

since

adenyl-cyclase.

mediators

involve

AND BIOPHYSICAL RESEARCH COMMUNICATIONS

been

the

of this

systematically have

investi-

also

cells

had

inter-

be

presence

The

might

in

might

revealed been

the

of

order

the

of 10e7N or more. or

Whether ved

in

our

results in

taken

trol.

At

this

levels

are of

actively

In

correlation

Mutants

have

normal be

these

coli,

we

not

find and

been

mention with

the

where

AMP

organism,

an this

type

of

a number

of

the

that

view

its

effects the

con-

recent

intra-

"controller"

unique

observations,

failed

to

cyclic

find AMP

and

during a consistent

intracellular

in our

isolated of

insensitive Wayne

AMP concentration repression

in

cyclic

have

of

cata-

been

most

appear

following

clearly

any

significant

concentration

and degree

("1

pattern

totally

Recently

do

to

invol-

of

in

involvement

systems

Among

repression.

completely

-

those

effect

synthesis

constitute

intracellular

between

2.

("I

an

to reconcile

in

Escherichia

catabolite

to

normal

actually

in BaciZZus megaterim,

observe

to

relevant

AMP

is

significant.

1.

out

its

compound

enzyme

difficult

investigated.

particularly

that

seems

even

other

repression

upon

cyclic

repression,

bolite

fact

of

it

some

catabolite

titrol

proof

as

which

cellular

in

point,

observations

of the

or

V&Y

be

they

control

illustrate

cannot

of

cyclic

normal

the

(whether

not

repression

catabolite to

Rosen

the [41

transient

addition

of

repression AMP

in between

levels.

351

which

which, cyclic

a decrease

found

correlation cyclic

laboratory,

exhibit

a

however,

turns

AMP.

in cellular

Escherichia permanent

cyclic eozi. [catabolitel

But

Vol. 57, No. 2, 1974

3. tants

have

while

the

Also

AMP

and

proline

other

of

oxidase

sum

if

of

these

of

is

strongly

both

to,

rather

that

the gift

the

(5). synthesis

of

of

suggests

mediated, of

AMP

in

cyclic

the

many

controls

view

organisms

mediators

involving

itself.

tvish to thank Dr. Jean-Pad

suggestions and for

galactokinase,

independent

strongly

a complex

cyclic

of

mu-

(61.

may be normally

addition

negative

repressed

quite

starvatibn

by

cyclase

levels

observed

are

nitrogen

systems,

Aknowledgements - I

teful

it

RESEARCH COMMtiNlCATiONS

and

normal is

observations

most

or in

than,

synthesize

histidase

repression not

negative

6-galactosidase

and

conditions

catabolite many

which

CAP

KlebsieZZa aerogenes,

In

The that

isolated

formation

under

AND BIOPHYYCAL

Escherichia coli.

in

been

4. both

BIOCHEMICAL

Aubert for many heZpful

of the strain used in this work.

to Dr. Jacques Monod for stimulating

I am gra-

discussions and criticism

of

the manuscript. This work was supported by grarts from the la Recherche Recherche

Scientifique

et

the "Centre

Technique”,

and the

Scientifique”

“Delegation

“National

Generale

National

Institutes

of

de

a la

Health”.

REFERENCES

1.

Setlow.

P.,

2.

Aubert,

J.-P.,

tit&

Biochem.

(19651.

3.

Ullmann,

A.,

4.

Wayne,

5.

Rothman-Denes,

6.

Prival,

and

cellulaires

545-551

1040

Biophys.

P.K.,

Res.

J.

Millet,

Comm.,

-52,

and

F.,

Rosen, L.B.,

and O.N.,

Hesse,

J.,

Nonod, in

[19731.

"M&anismes de re'gulation

in

chez Zes microorganismes",

Jacob,

365

J.

ed.

Vol.

by

C.N.R.S..

Viol..

-32.

des actiParis,

1 (1968).

press.

J.E.,

and

Epstein,

W.,

J. Bact..

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(19731.

M.J.,

and

Magasanik,

6..

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6266

(1971).