Table 1.

Phenotypes associated with C. burnetii T4SS substrates

Cbu no.	Screen and/or reference no.a	LP TEM (%)b	Ectopic localizationc	Yeast toxicityd	SEAPe	Transposon mutant/replication defectf
Cbu0012g	E-block	1	No pattern	No phenotype	No phenotype	Yes/slight defecth
Cbu0041 (cirA)	18	70	Punctate	Toxic	No phenotype	Yes/no intracellular replication
Cbu0069	PmrA	1	No pattern	No phenotype	No phenotype
Cbu0080	E-block; 12	70	No pattern	No phenotype	No phenotype
Cbu0113	E-block	7	No pattern	No phenotype	No phenotype
Cbu0129	18	70	Nucleic	No phenotype	No phenotype
Cbu0175	18	20	No pattern	No phenotype	No phenotype
Cbu0183	PmrA	20	No pattern	No phenotype	No phenotype
Cbu0201g	PmrA	1	No pattern	No phenotype	No phenotype	Yes/no defect
Cbu0212	E-block	10	No pattern	No phenotype	No phenotype
Cbu0270g	E-block	1	No pattern	No phenotype	No phenotype
Cbu0295g	E-block; 12	2	No pattern	No phenotype	No phenotype
Cbu0344	E-block	5	No pattern	No phenotype	No phenotype	Yes/no defect
Cbu0372	E-block	20	Endoplasmic reticulum	No phenotype	No phenotype	Yes/no defect
Cbu0375	E-block	35	No pattern	No phenotype	No phenotype
Cbu0376	E-block	7	No pattern	No phenotype	No phenotype
Cbu0388	PmrA	40	Nucleic	No phenotype	No phenotype	Yes/no intracellular replicationh
Cbu0393g	PmrA	1	Nucleic	No phenotype	No phenotype
Cbu0410	18	45	ND	No phenotype	No phenotype
Cbu0414	18	45	No pattern	No phenotype	No phenotype
Cbu0425 (cirB)	E-block; 12	90	Cytoplasmic	No phenotype	No phenotype	Yes/no intracellular replication
Cbu0469g	E-block	1	No pattern	No phenotype	No phenotype
Cbu0513	E-block	20	Cytoplasmic	No phenotype	No phenotype
Cbu0590	E-block	70	No pattern	No phenotype	No phenotype
Cbu0606	E-block	2	Cytoplasmic	Toxic	No phenotype
Cbu0637g	PmrA	1	No pattern	No phenotype	No phenotype
Cbu0665	PmrA	3	Cytoplasmic	No phenotype	No phenotype
Cbu0773	E-block	5	No pattern	No phenotype	No phenotype
Cbu0794	18	80	Nucleic	No phenotype	No phenotype
Cbu0801	18	10	No pattern	No phenotype	No phenotype
Cbu0814	18	10	No pattern	No phenotype	No phenotype
Cbu0881	18	80	ND	No phenotype	No phenotype
Cbu0885	E-block	1	Cytoplasmic	Toxic	No phenotype
Cbu0937 (cirC)	18	2	Cytoplasmic	No phenotype	No phenotype	Yes/no intracellular replication
Cbu1045	18	40	No pattern	No phenotype	No phenotype
Cbu1079	E-block	5	No pattern	No phenotype	No phenotype
Cbu1102	E-block	70	No pattern	No phenotype	No phenotype
Cbu1110	PmrA	40	No pattern	No phenotype	No phenotype
Cbu1150g	E-block	1	No pattern	No phenotype	No phenotype
Cbu1198	PmrA	75	No pattern	No phenotype	No phenotype	Yes/slight defecth
Cbu1217	18	10	No pattern	No phenotype	No phenotype
Cbu1314	18	2	Nucleic	No phenotype	No phenotype
Cbu1379g	18	1	No pattern	No phenotype	No phenotype
Cbu1406	18	40	Cytoplasmic	No phenotype	No phenotype
Cbu1425g	18	1	Mitochondrial	No phenotype	No phenotype
Cbu1434g	E-block	1	Cytoplasmic	No phenotype	No phenotype
Cbu1457	18	2	ND	No phenotype	No phenotype	Yes/slight defecth
Cbu1460	18	40	ND	No phenotype	No phenotype
Cbu1461	18	2	Cytoplasmic	No phenotype	No phenotype
Cbu1524	12, 18	10	Nucleic	Toxic	No phenotype
Cbu1525	E-block; 12		Cytoplasmic	No phenotype	No phenotype
Cbu1543	18	25	Punctate	No phenotype	No phenotype
Cbu1556	18	50	Cytoplasmic	No phenotype	0.23 ± 0.01	Yes/no defect
Cbu1566	E-block	5	No pattern	No phenotype	No phenotype
Cbu1569	18	90	No pattern	No phenotype	No phenotype	Yes/no defect
Cbu1576	PmrA	2	Endoplasmic reticulum	No phenotype	No phenotype
Cbu1594	Homolog	5	No pattern	No phenotype	No phenotype
Cbu1599	18	10	ND	No phenotype	No phenotype
Cbu1607	E-block	5	No pattern	No phenotype	No phenotype
Cbu1620	E-block	90	No pattern	No phenotype	No phenotype
Cbu1636	18	1	Punctate	No phenotype	No phenotype	Yes/no defect
Cbu1639	PmrA	4	No pattern	No phenotype	No phenotype	Yes/no defect
Cbu1665g	E-block	1	No pattern	No phenotype	No phenotype	Yes/no defect
Cbu1677	E-block	5	No pattern	No phenotype	No phenotype
Cbu1685	PmrA	1	No pattern	No phenotype	No phenotype
Cbu1751	18	50	No pattern	No phenotype	No phenotype
Cbu1754	E-block	2	No pattern	No phenotype	No phenotype
Cbu1769	18	1	No pattern	No phenotype	No phenotype
Cbu1789g	E-block	1	No pattern	No phenotype	No phenotype
Cbu1790g	E-block	1	Cytoplasmic	No phenotype	No phenotype
Cbu1823	12, 18	50	No pattern	No phenotype	No phenotype
Cbu1825	12, 18	40	Mitochondrial	No phenotype	1.70 ± 0.01
Cbu2007	E-block	90	Cytoplasmic	No phenotype	No phenotype	Yes/no defect
Cbu2013	PmrA	50	Cytoplasmic	No phenotype	No phenotype	Yes/slight defecth
Cbu2016g	E-block	1	No pattern	Toxic	No phenotype	Yes/no defect
Cbu2028g	E-block	1	No pattern	No phenotype	No phenotype
Cbu2052 (cirD)	12, 18	90	Cytoplasmic	Toxic	No phenotype	Yes/no intracellular replication
Cbu2059 (cirE)	E-block; 12	40	Cytoplasmic	No phenotype	No phenotype	Yes/no intracellular replication
Cbu2064g	E-block; 12	1	No pattern	No phenotype	No phenotype
Cbu2076g	E-block	1	No pattern	No phenotype	No phenotype
Cbu2078	12	10	ND	No phenotype	No phenotype
CbuA0014	12, 20	10	No pattern	No phenotype	No phenotype
CbuA0015	E-block, 20	90	No pattern	No phenotype	No phenotype
CbuA0019g	E-block	1	Cytoplasmic	No phenotype	1.36 ± 0.04
CbuA0020g	E-block	1	Mitochondrial	No phenotype	No phenotype

^aBioinformatic screen or reference used in identification of T4SS substrates.

^bTranslocation efficiency of C. burnetii T4SS substrates by L. pneumophilia.

^cSubcellular localization of GFP-tagged substrates ectopically expressed in HeLa cells (Fig. 2). ND, not determined.

^dHeterologous expression in yeast diminishes replication (Fig. 3).

^eExternal/internal SEAP ratio (Fig. 4). Numbers in bold represent values representing significantly (P < 0.01) diminished growth compared to vector and wild-type results.

^fTransposon mutants were isolated using rescue cloning, and growth was monitored in J774 and HeLa cells (Fig. 5).

^gPutative T4SS substrate with low translocation in Legionella; translocation not confirmed in the natural C. burnetii host.

^hGrowth defect in single mutant.