Figure S1. in vivo 30S and 50S ribosome assembly maps adapted from (16). (A) 30S ... Ribosomes of the indicated strains were isolated by sucrose cushion ...
A
5'
3' 16S RNA
S17
S4
S8
S16
S12
21S
S15
S20
S6:S18
S5
S11
S7 S9
S19
S10
S14
S3
S2
S21
B
S13
5'
3' 23S RNA
L24
32S
L20
L21
L22
43S
L23
L17
L3 L13
L34
L29
L19
L32
L5
L2
L14
L6 L33
L28
L30
L10
L18
5S
L11
L9
L16
L7/12 L35
L1
L15
L4
L27
L25 L31
L36
Figure S1. in vivo 30S and 50S ribosome assembly maps adapted from (16). (A) 30S assembly map reflecting order and interdependency of r-protein interaction with the nascent small ribosomal subunit. Early interacting r-proteins are shaded in dark gray, late interacting ones in light gray. Boxed proteins are contained in the 21S precursor of the 30S subunit. S2 is circled in red. (B) 50S assembly map reflecting order and interdependency of r-protein interaction with the nascent large ribosomal subunit. Early interacting r-proteins are shaded in dark gray, late interacting ones in light gray. Proteins contained in the 32S or in the 43S precursor of the large ribosomal subunit are boxed or cycled in black, respectively. L19 is circled in green.
.
S2-mCherry L19-EGFP
*
S2
28 17 10 *
MC rg
MC 41 0 MC 0 r MC g 75 63 48 35
*
L19
Figure S2. Immunoblot analysis of the supernatant after sucrose cushion centrifugation. Ribosomes of the indicated strains were isolated by sucrose cushion centrifugation. The protein content of the supernatant was TCA precipitated and analyzed by SDS-PAGE and subsequent immunoblotting using S2 and L19 specific antisera. Asterisks denote unspecific protein bands.
+IPTG
-IPTG
MC4100 MCΔsQ MCΔlC MCrg MCrgΔsQ MCrgΔlC Figure S3. Viability test of rpsQ and rplC conditional gene knock-out. Serial dilutions of cultures of the indicated strains were spotted on LB agar plates containing IPTG (left panel), or not (right panel). Cells were incubated at 37°C until visible colonies had formed.
A
debries
none
de
br ie
s
70S
2
B
10
20
30S
30
50S
40
50
60
70
23S rRNA 16S rRNA (fraction)
50
60
70
23S rRNA 16S rRNA (fraction)
70
23S rRNA 16S rRNA (fraction)
debries
de
br ie
s
Cam
2
C
10
20
30
40
debries
de br ie
s
Ery
2
10
20
30
40
50
60
Figure S4: A254 detection and fluorescence determination of scrose density fractions Sucrose density gradient (10-25%) centrifugation profiles from (A) control cells with no antibiotic (none), (B) chloramphenicol (Cam) and (C) erythromycin (Ery) treated cells. A254 profiles and fluorescence bar charts were superimposed and sucrose fractions were analyzed for presence of 16S and 23S rRNA by agorse gelelectrophoresis. Gray circles: low-intermediate ammounts of rRNA, black circles: high amounts of rRNA
