Table 1.

Strains and plasmids used in this study

Strain or plasmidRelevant propertiesSource or reference
Strains
    E. coli
        XL1-BlueendA1 gyrA96(Nalr) thi-1 recA1 relA1 lac glnV44 F′[::Tn10 proAB+ lacIq Δ(lacZ)M15] hsdR17(rK+ mK)Stratagene
        BL21(DE3)(pREP4-groESL)F ompT gal dcm lon hsdSB(rB mB) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5])2
    L. pneumophila
    CIP 108286Virulent L. pneumophila serogroup 1, strain Lens8
        ΔdotA mutantLens lpl2613::Km23
        ΔlegK1 mutantLens lpl1545::KmThis study
        ΔlegK2 mutantLens lpl2066::KmThis study
        ΔlegK3 mutantLens lpl2481::KmThis study
        ΔlegK4 mutantLens lpl0262::KmThis study
        ΔlegK5 mutantLens lpl2476::KmThis study
    A. castellaniiEnvironmental isolateP. Pernin, Faculty of Pharmacy, Université Lyon 1
    D. discoideum
        DBS0302388Wild-type DH1 strain12
        DBS0236184[act15]:cnxA:GFP55
        Dd03[act15]:legK2(K112M):c-mycThis study
        J774A.1ATCC TIB-67
Plasmids
    pGEX-6P-3E. coli expression vector69
    pGEX-legK1lpl1545 inserted in BamHI-SalI of pGEX-6P-3 for GST-LegK1 overproductionThis study
    pGEX-legK2lpl2066 inserted in BamHI-SalI of pGEX-6P-3 for GST-LegK2 overproductionThis study
    pGEX-legK3lpl2481 inserted in BamHI-SalI of pGEX-6P-3 for GST-LegK3 overproductionThis study
    pQE30E. coli expression vectorQiagen
    pQE30-legK4lpl0262 inserted in BamHI-SalI of pQE30 for 6His-LegK4 overproductionThis study
    pQE30-legK5lpl2476 inserted in PstI-SacI of pQE30 for 6His-LegK5 overproductionThis study
    pKD13Used for amplification of Km14
    pAV695sacB, OriT OriV, cml (pCDP05 with a deletion of a 4.3-kb fragment)Modified from reference 60
    p695-legK1::KmpAV695 derivative carrying lpl1545 interrupted by KmThis study
    p695-legK2::KmpAV695 derivative carrying lpl2066 interrupted by KmThis study
    p695-legK3::KmpAV695 derivative carrying lpl2481 interrupted by KmThis study
    p695-legK4::KmpAV695 derivative carrying lpl0262 interrupted by KmThis study
    p695-legK5::KmpAV695 derivative carrying lpl2476 interrupted by KmThis study
    pXDC50Legionella expression vector carrying mCherryX. Charpentier
    plegK1pXDC50 derivative Legionella expression vector carrying mCherry; expression of legK1 under the control of legK2 promoter (400 bp)This study
    plegK2pXDC50 derivative Legionella expression vector carrying mCherry; expression of legK2 under its promoter (400 bp)This study
    plegK2(K112M)pXDC50 derivative Legionella expression vector carrying mCherry; expression of legK2(K112M) under its promoter (400 bp)This study
    plegK3pXDC50 derivative Legionella expression vector carrying mCherry; expression of legK3 under the control of legK2 promoter (400 bp)This study
    plegK4pXDC50 derivative Legionella expression vector carrying mCherry; expression of legK4 under the control of legK2 promoter (400 bp)This study
    plegK5pXDC50 derivative Legionella expression vector carrying mCherry; expression of legK5 under the control of legK2 promoter (400 bp)This study
    p3041pXDC50 derivative Legionella expression vector carrying mCherry and gentamicin resistance geneThis study
    pEP46D. discoideum expression vector carrying legK2(K112M)-c-myc under the control of the actin promoterThis study
    pEP73Legionella expression vector carrying gfp-legK2; production of the GFP-LegK2 hybrid proteinThis study