TABLE 2.

Mutants selected in 8325-4 as Kanr SCVs and then screened for FA resistance

Strain and stock (SCV)Identified mutationaMIC (μg/ml) of:Doubling time (min)bAuxotrophycnd
fusA (EF-G)rplF (L6)KANSTRFA
8325-4
    AH0011.530.12524
FusA-SCV
    AH366 (SCV-38)Pro114HisNone12241277Hemin
    AH369 (SCV-41)Gln115LeuNone6124852Hemin
    AH364 (SCV-36)Asp434AsnNone82425664Hemin
    AH334 (SCV-6)Thr436IleNone1281924854Hemin + menadione
    AH336 (SCV-8)His438AsnNone1612865Hemin
    AH338 (SCV-10)Arg464CysNone4824267Hemin2
    AH340 (SCV-12)Gly617AspNone1224452Hemin7
    AH332 (SCV-4)Ala655GluNone12121268Hemin2
    AH329 (SCV-1)Arg659CysNone12241263Hemin5
    AH331 (SCV-3)Arg659HisNone1624849Hemin3
    AH347 (SCV-19)Arg659SerNone12321291Hemin
    AH367 (SCV-39)Gly664SerNone12161246Hemin2
FusE
    AH380 (SCV-52)None−1 FS at nt 158424652Hemin
    AH352 (SCV-24)None−1 FS at nt 106624453Hemin
    AH377 (SCV-49)None+1 FS at nt 23624449Hemin
    AH348 (SCV-20)NoneTGA stop at nt 244816862Hemin
    AH370 (SCV-42)eNoneTAA stop at nt 229824846Hemin
    AH346 (SCV-18)None−1 FS at nt 238824877Hemin
    AH333 (SCV-5)NoneTAA stop at nt 2498241256Hemin
    AH392 (SCV-64)None−1 FS at nt 3831216457Hemin
    AH379 (SCV-51)NoneTAA stop at nt 4181216645Hemin2
    AH349 (SCV-21)None−1 FS at nt 4041664467Hemin
    AH374 (SCV-46)None−5 FS at nt 236-2403296859Hemin
    AH358 (SCV-30)fNone−1 FS at nt 423486412100Hemin
    AH339 (SCV-11)None−1 FS at nt 144641286120Hemin
    AH337 (SCV-9)eNone−1 FS at nt 2209625612130Hemin
    AH357 (SCV-29)NoneTAA stop at nt 301962561279Hemin
    AH384 (SCV-56)gNone−1 FS at nt 22112825624140Hemin
    AH378 (SCV-50)e,gNone−1 FS at nt 221162568200Menadione
    AH351 (SCV-23)None−4 FS at nt 45-486425616180Menadione
    AH360 (SCV-32)NoneATA start at nt 39619224140Menadione
    AH383 (SCV-55)NoneTAA stop at nt 1399619216200Menadione
    AH362 (SCV-34)NoneTAA stop at nt 28212825612170Menadione
    AH330 (SCV-2)fNone−1 FS at nt 4231289612200Menadione
    AH390 (SCV-62)None+1 FS at nt 842566416170Menadione
    AH386 (SCV-58)eNone−1 FS at nt 902569624180Menadione
  • a None, wild type. Underlined letters represent the mutated residue. FS, frameshift mutation.

  • b Doubling time (generation time) in minutes during exponential growth in LB.

  • c Auxotrophy tested for hemin, menadione, thymine, and thymidine. −, nonauxotrophic.

  • d Number of independent isolates with the same genotype and phenotype.

  • e SCV-42, -9, -50, and -58 were sequenced for all ribosomal proteins and rRNA operons to determine whether any other mutations in the ribosome could distinguish between the hemin- and menadione-auxotrophic classes.

  • f One of a pair of mutants with the same mutation in rplF but different phenotypes.

  • g One of a pair of mutants with the same mutation in rplF but different phenotypes.