Supplemental Table S1: Plasmids
Plasmid / GenotypepDG268 / amyE::lacZ cat amp (Antoniewski et al., 1990)
pDG364 / amyE::cat amp (Guérout-Fleury et al., 1996)
pDG1664 / thrC::mls amp (Guérout-Fleury et al., 1996)
pDP294 / amyE::PyloC-lacZ cat amp
pDP296 / amyE::PremB-lacZ cat amp
pJW7 / amyE::PremA-remA cat amp
pJW8 / amyE::PyloC-remA cat amp
pJW10 / amyE::PremB-remB cat amp
pJW11 / thrC::PremA-remA mls amp
pJW13 / amyE::PrecF-remB cat amp
pJW18 / thrC::PrecF-remB mls amp
pJW21 / amyE::PrecF-lacZ cat amp
pJW23 / amyE::PremA-lacZ cat amp
pMarA / TnYLB-1 amp mls oriBsTs (Le Breton et al., 2006)
Supplemental Table S2: Primers
766 / GGGAATCATTTGAAGGTTGGT
749 / TTAGAGTTATTAATGGAATTGCTGATNNNNNNNNNNN
884 / AGCGTTGATGGCATTGATGACGA
885 / TGTTGATAGTGCGTTTACGACCGA
906 / TTCGTGATATATTAGCTAGTGGCG
907 / AACATAATCGACTGGCGTGCCG
939 / AGGAGGAATTCATCCATGCAGGCATCTCAAGAGG
1053 / AGGAGGAATTCGGCTCTTAGCGAAAGACTGC
1054 / CTCCTGGATCCAGAGAACGATTAATAACCCTCTTTCTT
1055 / AGGAGGAATTCGGAAGGAGACGTCATGAAGCGC
1056 / CTCCTCTCGAGTCATAATAAAAGGGCAATAGACACGC
1057 / AGGAGCTCGAGACTAGGCTAAACTAGAGACGTCTATT
1163 / AGGAGGAATTCCCAGTATGTCGGGGCCCTCAA
1164 / CTCCTGGATCCCTAAGAATCTATTTCAAACATAAATTG
1189 / AGGAGGAATTCCAACAACGTTGTAAAACTGTCCGCA
1190 / CTCCTCTCGAGTTTAATTGACGACTTGAAATGAAC
1191 / AGGAGCTCGAGGCGTTAGTGAAGTGAAGAAATG
1442 / CTCCTGGATCCCAATTCTTCCCTGTTCTCATAATAAAAGG
1443 / CTCCTGGATCCTTTCTTCACTTCACTAACGCACCA
Supplemental Table S3: β-galactosidase activity in LB medium1
Wild type / 17 ± 4.8 (DS1882) / 34.1 ± 13.1 (DS3352)
sinR / 150 ± 18.2 (DS2609) / 223 ± 10.5 (DS3391)
sinR remA / 4.1 ± 0.6 (DS2911) / 7.1 ± 3.0 (DS3374)
sinR remA PremA-remA / 25.7 ± 11.2 (DS3744) / 48.1 ± 2.1 (DS3745)
sinR remA PyloC-remA / 130 ± 11 (DS4260) / 213 ± 46 (DS4261)
sinR remB / 46.4 ± 8.6 (DS2617) / 27.8 ± 12.4 (DS3390)
sinR remB PrecF-remB / 63.5 ± 8.6 (DS4051) / 64.5 ± 28.7 (DS4052)
1All cultures were grown to 1.5 OD600 in LB medium. All values are the average of three replicas (Miller units ± standard deviation). All strains contain an epsH::tet allele to prevent cell clumping. This table contains the raw data for Figure 3a. Strains used to generate the data are indicated in parentheses.
Supplemental Table S4: β-galactosidase activity in MSgg medium1
Wild type / 45.6 ± 7.2 (DS1882) / 66.6 ± 12.2 (DS3352)
remA / 2.3 ± 1.5 (DS2913) / 1.2 ± 0.3 (DS3351)
remB / 6.3 ± 2.3 (DS2912) / 6.0 ± 3.1 (DS3407)
1All cultures were grown to 1.0 OD600 in MSgg medium. All values are the average of three replicas (Miller units ± standard deviation). All strains contain an epsH::tet allele to prevent cell clumping. This table contains the raw data for Figure 3b. Strains used to generate the data are indicated in parentheses.
SUPPLEMENTAL FIGURE LEGENDS
Figure S1: RemA and RemB proteins encoded by suppressor of sinR (sor) alleles. The predicted primary sequence of proteins encoded by two missense mutations in RemA
(DS1839 sor4, DS2326 sor31), one nonsense mutation in RemB (DS1837 sor2), and three frameshift mutation in RemB (DS2315 sor20, DS2319 sor24, DS2323 sor28). All suppressor mutations were independently isolated.
Figure S2: Mutations in remA and remB result in biofilm defects. Microtiter wells (6-well plate) within which cells have been grown in MSgg medium for 3 days at 25°C. The following strains were used to generate this figure: wild type (3610); remA (DS2679); remA PremA-remA (DS3330); remA PyloC-remA (DS3375); remB (DS2641), remB PremB-remB (DS3741); and remB PrecF-remB (DS4070).
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