Table 2.

Chronological appearance of mutations in tetracycline-resistant mutants

Locus IDa	Functiona	Mutationsb–d
		R6M1TC					R6M2TC
		1e	2f	3g	4h	5	1i	2j	3k	4
spr0187 (rpsJ)	30S ribosomal protein S10	G178T D60Y	G178T D60Y	G178T D60Y	G178T D60Y	C157T R53C Δ175–180	A169G K57E	A169G K57E	A169G K57E	A169G K57E
spr0219	phosphoglycerate mutase	WT	WT	WT	WT	T-52C	WT	WT	WT	WT
spr0291	phosphotransferase system IIA component	WT	WT	WT	WT	WT	C-101G	C-101G	C-101G	C-101G T-30C
spr0691	biotin synthase	WT	WT	WT	WT	WT	G356T A119D	G356T A119D	G356T A119D	G356T A119D
spr0822	agmatine deiminase	WT	WT	WT	WT	WT	WT	WT	WT	A95C Stop position 32
spr0825	hypothetical protein	G-61A	G-61A	G-61A	G-61A	G-61A	WT	WT	WT	WT
spr0960	positive transcriptional regulator MutR	WT	WT	WT	WT	A-151C	WT	WT	WT	WT
spr1262	transcriptional regulator Spx	WT	WT	T-11C	T-11C	T-11C	WT	WT	WT	WT
spr1377	cystathionine γ-synthase	WT	WT	WT	WT	WT	WT	WT	WT	T290 CA97V
spr1777	DNA-directed RNA polymerase subunit β	WT	WT	WT	WT	C1442T Syn	WT	WT	WT	WT
spr1885 (patB)	ABC transporter ATP-binding protein/permease	WT	WT	WT	WT	WT	WT	G794A G265E	G794A G265E	G794A G265E
spr1887 (patA)	ABC transporter ATP-binding protein/permease	G-35A	G-35A C883A R295S	G-35A G510A M170I C883A R295S	G-35A G510A M170I C883A R295S	G-35A G510A M170I C883A R295S	WT	WT	G-48A	G-48A G860A G287E
spr1991	glycerol kinase	WT	WT	WT	WT	WT	WT	WT	WT	ΔG position 75

^aID, identity. Nomenclature according to the genome annotation of S. pneumoniae R6.

^bWhen mutations are within coding regions, the change in amino acids is also indicated in italics.

^cIn non-coding sequence, the number preceded by a hyphen indicates the position upstream of the ATG.

^dMutations in bold were further studied to determine their potential role in tetracycline resistance and mutations underlined are involved in drug resistance (tetracycline, ciprofloxacin, ethidium bromide).

^eOne clone was sequenced. Unique mutations: spr1138 (C443A).

^fOne clone was sequenced. Unique mutations: spr0428 (G250A); spr0857 (G1298T).

^gTwo clones were sequenced. Unique mutations: spr0313 (A195G); spr1032 (G1216T); spr1367 (C511A); spr1865 (G224A); spr2035 (G-72A).

^hTwo clones were sequenced. Unique mutations: spr0294 (T111C).

ⁱTwo clones were sequenced. Unique mutations: spr0661 (G896T); spr0797: A to T 458 bp downstream of the gene.

^jTwo clones were sequenced. Unique mutations: spr0661 (G896T/A); spr1250 (T999C).

^kTwo clones were sequenced. Unique mutations: spr0232 (G191A).