The PprA-PprB two-component system activates CupE, the first non-archetypal Pseudomonas aeruginosa chaperone-usher pathway system assembling fimbriae

A

cupE1

cupE2

4648

4649

csuA/csuB

B

cupE5

cupE3 cupE4 4650

4651

cupE6

4652

4653

Chaperone

Usher

Adhesin-like protein

csuC

csuD

csuE

csuA csuB

1-27

122

183

186 aa

SCPU

CupE1 COG5430 1-28

119

174

177 aa

SCPU

CupE2 COG5430 1-24

177

119

SCPU

CupE3

180 aa

COG5430 1-24

97

162

SCPU1

CupE6 p

257

312

SCPU2 COG5430

C 28 29 25

315 aa

A

PAO1

PAO1DcupE5

B

PAO1DcupE5/ attB::cupE5

PAO1

PAO1DcupE5

PAO1DcupE5/ attB::cupE5

17

CupE1 DsbA

25

2 ,5

OD570nm

2 ,0

1 ,5

1 ,0

0 ,5

0 ,0

PAO1 PAO1 PAO1 PAO1 PAO1 DcupE5 DcupE5/ DcupE1 DcupE2 attB::cupE5

C

PAO1 DcupE3

PAO1 DcupE6

PAO1DcupE5

PAO1

D

PAO1DcupE5/attB::cupE5

N

31

17

D

E 500

500

10

shaking conditions static conditions

1 200

OD att 600nm

b-galactosiidase activity (Milleer units)

300

-galactosidase activity b (Miller units)

400 400

300 200 100 0

100

0 100

200

300 Time (min)

400

0,1 500

CupE1

17

A

B 87 1.

8

94 0.

24

0. 4

06

12

0.

0.

0.

PprB-6His ( M)

10000

0

b-galactosidase activity of cupE-lacZ (Miller units)

12000

8000

* 6000 4000 2000 0

17

CupE1 PAO1DpprB pMMB67HE

PAO1DpprB pMMB67pprB

PAO1DpprB pMMB67pprB6His

cupE (150bp)

A

6000 5000

shaking conditions static conditions

b-galactosidase activity (Miller units)

4000 3000 2000 500

0

PAO1/ pMMB67HE

B

PAO1/ pMMB67HE

CupE1

PAO1/ pMMBpprB

PAO1/ pMMBpprB

PAO1DpprB/ PAO1DpprB/ pMMB67HE pMMBpprB

PAO1DpprB/ PAO1DpprB/ pMMB67HE pMMBpprB

17

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