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Supplemental Informationdocx - Genes & Development

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Supplemental Informationdocx - Genes & DevelopmentGenes,&,genes

SUPPLEMENTAL INFORMATION

    Dual function of the McaS small RNA in controlling biofilm formation

    1,42,3,411,52,5Mikkel Girke Jørgensen, Maureen K. Thomason, Johannes Havelund, Poul Valentin-Hansen and Gisela Storz

    1Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark. 2Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD USA.

    3Department of Biochemistry and Molecular & Cellular Biology, Georgetown University Medical Center, Washington, DC USA.

4Co-first authors

    5Co-corresponding authors Email valentin@bmb.sdu.dk or storzg@mail.nih.gov

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Table of Contents

    Supplemental Figure S1 Additional truncations of pgaA 5’ UTR. Related to Figure 1. 4

    Supplemental Figure S2 Evidence supporting conservation of McaS and CsrA binding sites. Related to Figure 2. 5

    Supplemental Figure S3 Evidence supporting the dual functionality of McaS mutants, which are defective in 6

     CsrA binding but are still able to regulate targets by base pairing. Related to Figure 2. Supplemental Figure S4 Evidence supporting the conclusion that lower expression of McaS mutants is not the 7

     primary reason for failure to regulate pgaA-lacZ. Related to Figure 2.

     Supplemental Figure S5 Evidence supporting the specificity of CsrA for McaS and not other sRNAs such as the 9

     RyhB sRNA. Related to Figure 3.

    Supplemental Figure S6 Evidence showing that McaS regulates other CsrA targets such as glgC-lacZ. Related to 10

     Figure 5.

    Supplemental Figure S7 Quantitation of McaS, CsrB and CsrC levels throughout growth. Related to Figure 7. 11 Supplemental Figure S8 Effects of mutations in non-conserved GGA sequences. Related to Figure 2 and Discussion. 12

    Supplemental Figure S9 Evidence supporting the dual binding of CsrA and Hfq to McaS. Related to Figure 7 14

     and Discussion.

    Supplemental Table S1 List of strains and plasmids used in this study 16

    Supplemental Table S2 List of oligonucleotides used in this study 20

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Supplemental Methods Detailed descriptions of strain and plasmid construction. 30

Supplemental References 33

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Supplemental Figure S1. Assays of PM1205 ΔabgR-ydaL derivatives with (A) pgaA-lacZ and (B) pgaA-lacZ fusions transformed 15567

    with the control vector, pBR-McaS and plasmids expressing the McaS-2 and McaS-3 mutant derivatives. β-galactosidase activities of

    the fusions were assayed with either 1 mM IPTG (black bars) or no IPTG (white bars). The average values from three independent

    assays are shown and error bars are standard deviations of those values.

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