. 2014 Jul;196(14):2563–2577. doi: 10.1128/JB.01596-14

TABLE 1.

Spontaneous PBr mutants

Strain^a	PB culture^b		PB binding^c	Locus^d	Predicted protein^d	Annotation^d	Mutation^e
Strain^a	Growth	SpeB	PB binding^c	Locus^d	Predicted protein^d	Annotation^d	Type	Change
WT	−	−	+	NA	NA	NA	NA	NA
PBr 1.1	+	+	+	04725	PstS	Phosphate ABC transporter	InDel	fs-aa6
PBr 2.1	+	+	+	04725	PstS		InDel	fs-aa6
PBr 3.1	+	+	+	03260	EbsA	Pore-forming protein	SNP	I95N
				04415	TopA	DNA topoisomerase I	InDel	fs-aa661
				04725	PstS		InDel	fs-aa6
PBr 3.2	+	+	+	00065	FtsH	Cell division protein	InDel	Δ293
				04720	PstC	Phosphate ABC transporter permease	SNP	Q126*
PBr 3.3	+	+	+	04725	PstS		InDel	fs-aa6
				09125	GdpP	GGDEF domain phosphodiesterase	InDel	fs-aa3
PBr 3.5	+	+	+	07215	FabT	Transcription repressor, fatty acid biosynthesis	SNP	S84L
				09125	GdpP		InDel	fs-aa218
				09180	YfmH	M16 family zinc protease	InDel	fs-aa140
PBr 3.6	+	+	+	02925	AgaS	Galactosamine-6P isomerase	SNP	G72D
				07145	ManN	Mannose-specific PTS IID protein	SNP	I75F
				07215	FabT		SNP	*145L
				09125	GdpP		InDel	fs-aa76
PBr 3.7	+	+	+	00065	FtsH		SNP	V291I
				04725	PstS		InDel	fs-aa6
PBr 4.1	+	−	+	03620	ClpX	Clp protease ATP-binding subunit	SNP	Q311*
PBr 4.4	+	+	+	04725	PstS		InDel	fs-aa6
				09125	GdpP		InDel	Δ642–645
PBr 4.5	+	+	+	03645	DeoB	Phosphopentomutase	SNP	S87Y
				04725	PstS		InDel	fs-aa6
PBr 6.3	+	+	+	07145	ManN		SNP	G166D
PBr 6.7	+	+	+	00060	Hpt	Hypoxanthine-guanine phosphoribosyltransferase	InDel	fs-aa20
				09125	GdpP		InDel	fs-aa563
PBr 6.10	+	+	+	00055	TilS	tRNA^Ile-lysidine synthase	InDel	Δ428
				00060	Hpt		InDel	ΔRBS-53
				07135	ManL	Mannose-specific PTS IIAB protein	SNP	N284K
PBr 9.21	+	−	+	03620	ClpX		InDel	Δ352–356
				07215	FabT		SNP	H39N
PBr 10.6	+	+	+	07215	FabT		SNP	S84L
				07750	NA	Phage repressor protein	InDel	(−24)
				09125	GdpP		SNP	Q547*
PBr 10.9	+	+	+	04415	TopA		InDel	fs-aa27
				04715	PstA	Phosphate ABC transporter permease	InDel	fs-aa35
				06715	Gmk	Guanylate kinase	SNP	L131I
PBr 10.11	+	−	+	01235	NanH	N-Acetylneuraminate lyase	SNP	[V120V]
				03620	ClpX		InDel	fs-aa178
				07215	FabT		SNP	A56V
PBr 10.14	+	+	+	04545	GuaA	GMP synthase	SNP	A248D
				04725	PstS		InDel	fs-aa6
PBr 10.15	+	+	+	00055	TilS		SNP	G428E
				00060	Hpt		SNP	(−9) RBS
PBr 11.1	+	−	+	03260	EbsA		InDel	Δ21–25
				03620	ClpX		SNP	E263*
				04650	NupP	Guanosine ABC transporter permease	SNP	W209*
				07215	FabT		SNP	(−9) RBS
PBr 11.2	+	+	+	00055	TilS		SNP	*429R
				00060	Hpt		SNP	(−7) RBS
				04415	TopA		SNP	R646*
PBr 11.3	+	+	+	05585	PtsI	PEP^f-protein phosphotransferase	SNP	A510V
				08845	LuxR	Transcriptional regulator, LuxR family	SNP	D693Y
				09125	GdpP		SNP	A574D
PBr 11.5	+	+	+	07135	ManL		SNP	N284K
				08035	Fba	Fructose-bisphosphate aldolase	SNP	A6V
PBr 11.6	+	+	+	04415	TopA		SNP	V29D
				05585	PtsI		SNP	D164Y

WT is HSC5. Mutants whose designations begin with different numbers were isolated from independent experiments.

Ability (+) or inability (−) to form colonies (growth) and express SpeB protease activity (SpeB) when grown on protease indicator plates supplemented with 400 μg/ml PB.

Focal (+) or undetectable (−) membrane binding when stained with 5 μM dansyl-PB.

Genetic loci, the predicted proteins that an ORF encodes, and annotations are based on the genome of HSC5 (16). Loci follow the format L897_xxxxx, where xxxxx are numbered. NA, not applicable. Annotations are listed only at first instance. In subsequent examples, the field is left blank.

InDel, insertion/deletion; SNP, single nucleotide polymorphism; “fs-aa” followed by numeral, frameshift at the indicated codon; *, stop codon; number in parentheses, distance upstream of start codon; RBS, ribosome binding site; data in brackets, synonymous mutation; Δ, deletion. Note that tilS and hpt mutations result from a single alteration since the 3′ end of tilS overlaps the RBS of hpt.

PEP, phosphoenolpyruvate.