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ESX/Type VII secretion systems and their role

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Available online at www.sciencedirect.com

ESX/type VII secretion systems and their role in hostpathogen

interaction
Roxane Simeone1, Daria Bottai1,2 and Roland Brosch1
The ESX-1 system is responsible for the secretion of the
prototypic ESX proteins, namely the 6 kDa early secreted
antigenic target (ESAT-6) and the 10 kDa culture filtrate protein
(CFP-10). These two proteins, which form a 1:1 heterodimeric
complex, are among the most important proteins of
Mycobacterium tuberculosis involved in hostpathogen
interaction. They induce a strong T cell mediated immune
response, are apparently involved in membrane and/or host
cell lysis and represent key virulence factors. There are four
other paralogous ESX systems in M. tuberculosis, some of
which are essential for in vitro growth. ESX systems also exist in
many other actinobacteria and Gram-positive bacteria, and
have recently been suggested to be named type VII secretion
systems.
Addresses
1
Institut Pasteur, UP Pathogenomique Mycobacterienne Integree, 25
Rue du Dr. Roux, 75724 Paris, France
2
University of Pisa, Dipartimento di Patologia Sperimentale,
Biotecnologie Mediche, Infettivologia ed Epidemiologia, Pisa, Italy
Corresponding author: Brosch, Roland (roland.brosch@pasteur.fr)

Current Opinion in Microbiology 2009, 12:410

This review comes from a themed issue on
Host-microbe interactions: Bacteria
Edited by Brendan Kenny and Raphael Valdivia
Available online 18th January 2009
1369-5274/$ see front matter
# 2008 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.mib.2008.11.003

Introduction
Secreted proteins are among the most important molecules involved in hostpathogen interaction of Mycobacterium tuberculosis, the etiological agent of human
tuberculosis (TB). Proteins containing typical signal
sequences, such as the Ag85 T cell antigens get secreted
by the Sec mediated secretory pathway, and secretion of
other proteins may occur via the alternative Sec2 pathway
or the twin-arginine translocation (TAT) system. However, culture filtrate of M. tuberculosis also contains many
small, highly immunogenic proteins that lack classical
signal sequences. The 6 kDa early secreted antigenic
target (ESAT-6) [1] is the prototype of these proteins,
which are characterized by the amino acid (aa) motif TrpXaa-Gly (WXG), and a size of approximately 100 aa [2].
Multi-genome analyses predict the presence of WXG-100
proteins in a wide range of actinobacteria and GramCurrent Opinion in Microbiology 2009, 12:410

positive bacteria [2], where they are thought to be

secreted via novel secretion systems that were named
ESAT-6 systems (ESX-1-ESX-5) [3], or more recently
type VII secretion systems (T7S system) [4]. In mycobacteria these clusters typically also encode PE and PPE
proteins characterized by their N-terminal motifs prolineglutamic acid (PE) and proline-proline-glutamic acid
(PPE) [5].
ESX-1 of M. tuberculosis and related tubercle bacilli is
the ESX/T7S system that has probably attracted the
most attention in recent years. Located in the region of
difference 1(RD1), ESX-1 is deleted in the vaccine
strain Mycobacterium bovis BCG and other attenuated
members of the M. tuberculosis complex like the vole
bacillus Mycobacterium microti, suggesting that secretion
of ESAT-6 is involved in virulence of tubercle bacilli.
First experimental evidence confirming this hypothesis
was obtained by complementing BCG and M. microti,
with differently sized portions of the RD1 region from
M. tuberculosis [6,7]. Only strains that were complemented with the entire RD1 locus and its flanking regions
showed secretion of ESAT-6 whereas strains complemented with smaller constructs expressed ESAT-6 but
failed to secrete it (Figure 1). Recombinant BCG::RD1
and M. microti::RD1 strains with a fully restored ESX-1
secretion system grew more vigorously in severe combined immunodeficient mice and also persisted longer
in the organs of immunocompetent mice [7]. However,
they induced considerably less pathology than M. tuberculosis, suggesting that the loss of the ESX-1 system
from BCG was only one of several factors that contributed to the attenuation of the vaccine strain, as was
recently confirmed by comparative and functional
genomics [8].
In parallel, studies using transposon mutants and/or RD1
knockout strains of M. tuberculosis came to the same
conclusion that RD1 dependent ESAT-6 secretion was
essential for virulence of M. tuberculosis [912].
ESX-1 is conserved in several other mycobacteria, such as
Mycobacterium leprae and the fish pathogen Mycobacterium
marinum. Whereas work on ESX-1 in M. leprae has been
discouraged by the non-culturable status of the bacterium, the genomic resemblance of ESX-loci in M. marinum
and M. tuberculosis [13] inspired numerous studies that
use M. marinum as a model [14]. The only available data
on ESX cluster 5 (ESX-5) also come from M. marinum,
and revealed that ESX-5 is implicated in the secretion of
PE and PPE proteins [15,16].
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ESX/type VII secretion systems and their role in hostpathogen interaction Simeone, Bottai and Brosch 5

Figure 1

Diagram of the M. tuberculosis H37Rv genomic region showing the position of various genes, depicted as differently colored arrows. Blue color
indicates that genes are essential for expression of ESAT-6 and CFP-10, whereas green color indicates that genes are needed for secretion of ESAT-6
and CFP-10 (after references [6,23]). The figure also shows the RD1mic region absent from M. microti (black bracket) [6,7] and the M. tuberculosis
inserts delineated by blue or red lines contained in integrating cosmids RD1-2F9 and pe-ppe-esxB-esxA (pAP34) that were used for complementation
of M. microti via the attB site. The results of western blot analyses using anti-ESAT-6 and anti-ESAT-CFP-10 antibodies (Ab) are depicted in the upper
left portion of the figure and clearly show that complementation with cosmid 2F9-RD1 leads to secretion of ESAT-6 and CFP-10 into the supernatant,
while complementation with construct pe-ppe-esxB-esxA (pAP34) leads to expression but not secretion of the two antigens (after reference [6]). Note
that brackets showing RD1BCG (Rv3871Rv3879) were omitted for simplicity.

Surprisingly, an orthologous ESX-1 system is also present

in the avirulent saprophyte Mycobacterium smegmatis,
where it is involved in conjugation [17]. Finally it should
be mentioned that the more distantly related ESX/T7S
systems in Gram-positive bacteria, may or may not have a
role in pathogenicity. In Listeria monocytogenes, no effect
on virulence could be attributed to ESX [18], while in
Staphylococcus aureus, the two ESAT-6-like proteins EsxA
and EsxB were associated with abscess formation in a
murine infection model [19]. Although the structures of
EsxA and EsxB from S. aureus resemble those of ESAT-6
(EsxA) and the 10 kDa culture filtrate protein CFP-10
(EsxB) of M. tuberculosis that are known to form a 1:1
heterodimeric complex [20], the staphylococcal Esx
proteins form mainly homodimers [21]. Finally, an
ESX/T7S system was also identified in Bacillus antrax,
where EsxB is required for secretion of EsxW and inducwww.sciencedirect.com

tion of humoral immune responses in infected guinea pigs

[22].
Taken together, it is clear that ESX/T7S systems of
actinobacteria and Gram-positive bacteria are highly interesting scientific topics studied by many research groups
worldwide. In this review we will present some key aspects
on the functional characterization of these systems and
their secreted proteins. Owing to space constraints, however, we will focus on the ESX-1 system of M. tuberculosis,
which may serve as a paradigm for the other systems.

The ESX-1 secretion machine

The contribution of individual ESX-1 proteins to
secretion of ESAT-6/CFP-10 is well documented
[6,10,11,23]. Whereas genes rv3867, rv3873, rv3876,
rv3878 and rv3879 do not appear to be essential for
Current Opinion in Microbiology 2009, 12:410

6 Host-microbe interactions: Bacteria

ESAT-6 and CFP-10 secretion in tubercle bacilli, the

remaining genes encoded in the extended RD1 region are
involved in this process (Figure 1). Rv3868 resembles an
ATPase associated with various cellular activities (AAA)
that, unlike previous predictions, may not fulfil a general
chaperone-like function [24]. Three other proteins
encoded upstream and downstream of esxA, Rv3869,
Rv3870 and Rv3877, exhibit 1, 3 or 11 predicted transmembrane domains, respectively. Together with Rv3871,
a cytosolic component of the ESX-1 system, these proteins
are thought to form a membrane-bound ESX-1 secretory
complex, with protein export driven by ATP hydrolysis
[3,4]. Recently, a C-terminal signal sequence has been
identified that allows the unstructured C-terminus of CFP10 to be recognized by Rv3871 that itself interacts with the
membrane protein Rv3870. Point mutation in this signal
sequence abolished binding of CFP-10 to Rv3871 and
prevented secretion of ESAT-6 and CFP-10 [25]. As
Rv3870 and Rv3871 both show similarity to proteins of
the FtsK-SpoIIIE ATPase family, these proteins might
perform an essential part of the work necessary to secrete
ESX-1 substrates. This mechanism resembles type IV
secretion system in Gram-negative bacteria, where a membrane-bound SpoIIIE/FtsK-like ATPase recognizes an
unstructured C-terminal sequence and directs the secreted
substrate to the cytoplasmic membrane [26].
Apart from genes localized in the RD1 region, a second
gene cluster (rv3616c-rv3614c) situated elsewhere in the
genome of M. tuberculosis, was shown to contribute to the
secretion of ESAT-6 and CFP-10 [27,28]. Inactivation of
these genes in M. tuberculosis abolished the secretion of
ESAT-6 and CFP-10, in spite of a complete RD1 locus
[27,28].

Regulation of gene expression in ESX-1

Until very recently the regulation of the ESX-1 system was
unknown. As ESAT-6 and CFP-10 are found in supernatants of in vitro grown M. tuberculosis cultures, secretion of
these antigens does not seem to dependent on interaction
with host cells. This feature is different to mechanisms that
govern regulation of type III and type IV secretion systems
of Gram-negative bacteria, which are switched on upon
hostcell contact. These characteristics of ESAT-6
secretion may imply that ESX systems, although being
essential for the pathogenicity of certain virulent mycobacteria, may fulfil more general roles in the physiology of
the bacteria than strictly hostcell oriented functions.
Adaptation towards pathogenesis may have evolved via
the second genomic region involved in ESAT-6 secretion,
carrying rv3616c-rv3614c. Orthologues of these genes are
missing from M. smegmatis and it remains to be determined
whether some distantly related, structurally similar
proteins may intervene in ESX-1 secretion in M. smegmatis
[17]. As the M. tuberculosis rv3616c-rv3614c gene cluster is
clearly involved in the regulation of ESAT-6 secretion,
such interspecies differences may be of importance. One of
Current Opinion in Microbiology 2009, 12:410

the identified key players in this process is PhoP, the

response regulator of the two-component system PhoR/
PhoP, which is essential for virulence of M. tuberculosis [29].
The link between a functional PhoP protein and ESAT-6
secretion was recently demonstrated by complementation
of the attenuated M. tuberculosis strain H37Ra with a wildtype copy of gene phoP that restored ESAT-6 secretion and
partially increased the virulence of the recombinant
H37Ra::phoP strain [30]. M. tuberculosis H37Ra carries a
point mutation in its phoP gene that interferes with the
DNA binding capacities of PhoP [30,31,32] and apparently abrogates secretion of ESAT-6 via reduction of
rv3616c-rv3614c expression [30]. Previous transcriptome
analyses that compared wild-type M. tuberculosis H37Rv
with M. tuberculosis H37Ra or a phoP knockout mutant of M.
tuberculosis H37Rv, respectively identified at least 30
genes, including cluster rv3616c-rv3614c, that were significantly lower expressed in both PhoP inactivated strains
[33,34]. However, PhoP is not the only regulator that
influences rv3616c-rv3614c expression and thereby ESAT6 secretion. A transcription factor named EspR, encoded by
gene rv3849, binds to the rv3616c-rv3614c promoter and is
itself secreted from M. tuberculosis by the ESX-1 system
[35]. Whether or not EspR is itself controlled by the PhoR/
PhoP remains to be determined. Available transcriptome
data are not conclusive, as in some studies that used strains
with inactivated or mutated PhoP, the expression level of
EspR was low [34,36], while in others such downregulation was not observed [33].
Apart from ESX-1, transcriptome data show that expression of proteins belonging to the ESX-3 cluster is controlled by the iron-dependent regulator IdeR [37] and the
zinc uptake regulator Zur (previously named FurB) [38].
This suggests that ESX-3 might be involved in fundamental biological processes such as metal ion homeostasis
of mycobacteria, which is consistent with the essentiality
of ESX-3 for in vitro growth of M. tuberculosis [39] and the
conservation of orthologous ESX-3 systems in a wide
range of mycobacterial species.

The ESX-1/T7S system effector proteins

involved in hostpathogen interaction
Although several features related to pathogenicity were
found to be associated with ESX-1, the primary biological
function of this system in M. tuberculosis and other ESX-1
harboring bacteria still remains unclear. Reported ESX-1related effects in tubercle bacilli include the suppression
of proinflammatory responses [11], necrosis [40], apoptosis [41], membrano-lysis [42] and cytolysis [10,12]. One
of the most intriguing questions in this respect is the
nature of the ESX effector molecules. Several lines of
evidence suggest that ESAT-6 and CFP-10 are the key
effector molecules of the ESX-1 system. However, it has
also been suggested that ESAT-6 and CFP-10 together
with Rv3616c (EspA) could serve as building blocks of the
ESX-1 secretion machinery in the cell envelope [27]. In
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ESX/type VII secretion systems and their role in hostpathogen interaction Simeone, Bottai and Brosch 7

this case ESAT-6, CFP-10 and EspA may simply be

sloughed off into the supernatant by mechanical forces.
However, the fact that certain single amino acid changes in
ESAT-6 did not prevent secretion of the modified ESAT-6
molecules but caused attenuation of the recombinant M.
tuberculosis strains [43] argues in favor of ESAT-6 being a
secreted effector molecule (Figure 2). Furthermore, the
finding that the C-terminal signal sequence of CFP-10 is
sufficient to secrete yeast fusion proteins via the ESX
system argues for CFP-10 being a secreted protein
[25]. Among the different proposed functions for
ESAT-6 and ESX-1 in the present scientific literature,
the most frequently reported observations point towards a
function linked to lysis of cells and/or membranes. Indeed,
Guinn et al. have reported that ESAT-6 deletion mutants of
M. tuberculosis could multiply within macrophage-like cells
lines but were unable to spread to uninfected macrophages
[10]. It was also described that ESAT-6 deleted mutants

show reduced tissue invasiveness due to lacking cytolytic

activity [12], and it was shown that ESAT-6 and CFP-10
interacted in a pH-dependent fashion with liposomes that
contained membranes with biologically relevant phospholipids [42], (Figure 2). Furthermore, cryoelectron microscopy revealed that ESAT-6 destabilized and lysed
liposomes, whereas CFP-10 did not [42]. Thus, the biological activity of ESAT-6 might depend on the acidity of
the environment, which is an important factor inside
phagosomes. By contrast, a different study using circular
dichroism spectroscopy suggested that complexes formed
by ESAT-6 and CFP-10 and/or other ESX heterocomplex
couples were too stable to dissociate at lower pH [44]. A
possible explanation for this discrepancy might be that the
first study [42] used culture filtrates from tubercle bacilli,
which normally contain acetylated ESAT-6, whereas the
second study [44] used non-acetylated ESAT-6 like
proteins expressed in Escherichia coli. In previous blot

Figure 2

Panel (a) shows a model of the ESAT-6-CFP-10 complex, with CFP-10 in white and ESAT-6 in blue. The mutations and their effect are indicated by
color codes. Red, enhanced virulence abolished; orange, reduced, intermediate virulence; green, ESX-1-enhanced virulence retained (after reference
[43]). Panel (b) Presence of ESAT-6 in the top fraction of a liposome floatation assay indicates interaction between ESAT-6 and liposomes, which is not
observed when the same amount of ESAT-6 is supplied via culture filtrate of M. tuberculosis and/or BCG::RD1 wherein ESAT-6 is present as a 1:1
complex with CFP-10, hiding its hydrophobic interphase. The right side of panel (b) shows results from liposome floatation analysis using BCG::RD1derived culture filtrates in buffers with different pHs, suggesting that at lower pH the complex formed by ESAT-6 and CFP-10 may dissociate and allow
binding of ESAT-6 to liposomes [42]. Panel (c) shows the results from surface plasmon resonance that confirm the effect of acidic pH on the
dissociation of the CFP-10-ESAT-6 complex [42]. Please note that the ESAT-6 used in that experiment was derived from M. tuberculosis that usually
carries an N-terminal acetylation [45], which might influence the stability of the complex. Panel (d) shows images obtained by cryoelectronmicroscopy
showing that ESAT-6 destabilizes and lyses liposomes, a feature that is not observed for CFP-10 [42].
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Current Opinion in Microbiology 2009, 12:410

8 Host-microbe interactions: Bacteria

overlay assays it was observed that CFP-10 interacted with

non-acetylated ESAT-6, while binding with acetylated
ESAT-6 was less pronounced [45].
Together, these features appear important when put in
context with the recently described translocation of M.
tuberculosis from the phagosome into the host cell cytoplasm at later stages of infection [46]. While at days 4 to
7 after infection with M. tuberculosis more than one third of
the non-apoptotic human dendritic cells contained translocated bacteria, this effect was not observed when BCG
or M. tuberculosis ESX-1 transposon mutants were used for
infection [46]. As M. tuberculosis is generally considered
to remain in phagosomes that resist maturation and acidification [47], these novel data on the translocation of M.
tuberculosis are still controversial. However, a recent independent study using human macrophages, also reported
that 25% of the M. tuberculosis cells at day 5 post-infection
were not enclosed by a phagosomal membrane [48]. As
such, the ESX-1 system might provide M. tuberculosis with
the tools necessary for escaping from the phagosomal
compartment of professional phagocytic cells and/or
releasing ESAT-6 proteins to the cytoplasm, where they
gain access to the class I-processing machinery contained
in the proteasome. Such events would explain the strikingly higher recruitment and activation of CD8+ T cells
found in the lungs of mice that were aerosol infected with
M. tuberculosis or BCG::RD1 than uninfected or BCGchallenged mice [49]. Indeed, there is also more and more
evidence from immunologic studies that ESX-1 proteins
gain access to the cytoplasm of the host cell [50].
In conclusion, the ESX/T7S systems represent novel
secretion systems, some of which are clearly involved
in pathogenicity and hostcell interaction. Great progress
has been made in the past five years to characterize these
systems, but there are still many details of their biology to
be elucidated that might have important consequences
for the design of novel antituberculosis vaccines and
therapeutic agents.

Acknowledgements
We are grateful to our colleagues Marien de Jonge, Alexander Pym, Priscille
Brodin, Marjan Fretz, Wafa Frigui, Laleh Majlessi, Claude Leclerc, Felix
Romain, Caroline Demangel, Patrick England, Gerard Pehau-Arnaudet,
Michael Nilges and Stewart Cole who in recent years contributed
substantially to the studies that are reviewed in this article. This work was
supported by the European Union (contracts LHSP-CT-2005018923,
HEALTH-F3-2007-201762) and the Institut Pasteur.

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by PhoP. PLoS Pathog 2008, 4:e33.
This paper provides for the first time evidence that ESAT-6 secretion is
regulated in M. tuberculosis and finds a link between the well-known twocomponent virulence regulator PhoP and ESAT-6 secretion that involves
the rv3616c-rv3614c gene cluster. This link opens new perspectives to
elucidate ESX and virulence regulation of M. tuberculosis.
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31. Wang S, Engohang-Ndong J, Smith I: Structure of the

DNA-binding domain of the response regulator PhoP
from Mycobacterium tuberculosis. Biochemistry 2007,
46:14751-14761.
32. Lee JS, Krause R, Schreiber J, Mollenkopf HJ, Kowall J, Stein R,
Jeon BY, Kwak JY, Song MK, Patron JP et al.: Mutation in the
transcriptional regulator PhoP contributes to avirulence of
Mycobacterium tuberculosis H37Ra strain. Cell Host Microbe
2008, 3:97-103.
33. Walters SB, Dubnau E, Kolesnikova I, Laval F, Daffe M, Smith I:

The Mycobacterium tuberculosis PhoPR two-component
system regulates genes essential for virulence and
complex lipid biosynthesis. Mol Microbiol 2006,
60:312-330.
This analysis provides at least 44 candidate genes that are regulated by
the two-component regulator PhoP, some of which have recently been
shown to be involved in ESAT-6 secretion.
34. Gao Q, Kripke K, Arinc Z, Voskuil M, Small P: Comparative

expression studies of a complex phenotype: cord formation in
Mycobacterium tuberculosis. Tuberculosis (Edinb) 2004,
84:188-196.
The supplementary data of this paper provide the results of an extraordinary large range of transcriptome comparisons between the virulent
H37Rv and the attenuated H37Ra strains of M. tuberculosis.
35. Raghavan S, Manzanillo P, Chan K, Dovey C, Cox J: Secreted

transcription factor controls Mycobacterium tuberculosis
virulence. Nature 2008, 454:717-721.
This paper describes the regulation of ESAT-6 secretion via influencing
expression of gene cluster rv3616crv3614c by the protein EspR
(Rv3849), which is itself secreted by ESX-1.
36. Gonzalo-Asensio J, Mostowy S, Harders-Westerveen J,
Huygen K, Hernandez-Pando R, Thole J, Behr M, Gicquel B,
Martin C: PhoP: a missing piece in the intricate puzzle of
Mycobacterium tuberculosis virulence. PLoS ONE 2008,
3:e3496.
37. Rodriguez GM, Voskuil MI, Gold B, Schoolnik GK, Smith I: ideR, an
essential gene in Mycobacterium tuberculosis: role of
IdeR in iron-dependent gene expression, iron metabolism,
and oxidative stress response. Infect Immun 2002,
70:3371-3381.
38. Maciag A, Dainese E, Rodriguez GM, Milano A, Provvedi R,

Pasca MR, Smith I, Palu G, Riccardi G, Manganelli R: Global
analysis of the Mycobacterium tuberculosis Zur (FurB)
regulon. J Bacteriol 2007, 189:730-740.
First evidence that the highly conserved ESX-3 system is regulated by the
Zink-uptake regulator Zur (previously named FurB) and might therefore be
involved in metal ion homeostasis.
39. Sassetti CM, Boyd DH, Rubin EJ: Genes required for
mycobacterial growth defined by high density mutagenesis.
Mol Microbiol 2003, 48:77-84.
40. Junqueira-Kipnis AP, Basaraba RJ, Gruppo V, Palanisamy G,
Turner OC, Hsu T, Jacobs WR Jr, Fulton SA, Reba SM, Boom WH
et al.: Mycobacteria lacking the RD1 region do not induce
necrosis in the lungs of mice lacking interferon-gamma.
Immunology 2006, 119:224-231.
41. Derrick SC, Morris SL: The ESAT6 protein of Mycobacterium
tuberculosis induces apoptosis of macrophages by
activating caspase expression. Cell Microbiol 2007,
9:1547-1555.
42. de Jonge MI, Pehau-Arnaudet G, Fretz MM, Romain F, Bottai D,

Brodin P, Honore N, Marchal G, Jiskoot W, England P et al.: ESAT6 from Mycobacterium tuberculosis dissociates from its
putative chaperone CFP-10 under acidic conditions and
exhibits membrane-lysing activity. J Bacteriol 2007,
189:6028-6034.
This paper provides evidence that suggests that ESAT-6 and CFP-10
bind to biological membrane when they are not forming a complex, which
may dissociate under certain conditions. The results shown in the paper
also suggest that ESAT-6 has membrane-lysing activity.
43. Brodin P, de Jonge MI, Majlessi L, Leclerc C, Nilges M, Cole ST,
Brosch R: Functional analysis of early secreted antigenic
target-6, the dominant T-cell antigen of Mycobacterium
tuberculosis, reveals key residues involved in secretion,
Current Opinion in Microbiology 2009, 12:410

10 Host-microbe interactions: Bacteria

complex formation, virulence, and immunogenicity. J Biol

Chem 2005, 280:33953-33959.
44. Lightbody KL, Ilghari D, Waters LC, Carey G, Bailey MA,
Williamson RA, Renshaw PS, Carr MD: Molecular features
governing the stability and specificity of functional complex
formation by Mycobacterium tuberculosis CFP-10/ESAT-6
family proteins. J Biol Chem 2008, 283:17681-17690.
45. Okkels LM, Muller EC, Schmid M, Rosenkrands I, Kaufmann SH,
Andersen P, Jungblut PR: CFP10 discriminates between
nonacetylated and acetylated ESAT-6 of Mycobacterium
tuberculosis by differential interaction. Proteomics 2004,
4:2954-2960.
46. van der Wel N, Hava D, Houben D, Fluitsma D, van Zon M,
 Pierson J, Brenner MJPP: M. tuberculosis and M. leprae
translocate from the phagolysosome to the cytosol in myeloid
cells. Cell 2007, 129:1287-1298.
The work described in this paper shows evidence that at later stages of
infection one third to one half of the M. tuberculosis cells in professional
phagocytes leave the phagosomes and enter the host cell cytoplasm, a
feature that is not observed for ESX-1 lacking BCG and CFP-10/ESAT-6
mutants. This paper provides clear-cut cryoelectronmicroscopical
images of M. tuberculosis cells in the hostcell cytoplasm. The paper
is controversial as it tries to change a long lasting dogma that M.
tuberculosis resides and multiplies only in the phagosomal compartment.

Current Opinion in Microbiology 2009, 12:410

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47. Russell DG: Phagosomes, fatty acids and tuberculosis. Nat Cell
Biol 2003, 5:776-778.
48. Lee BY, Clemens DL, Horwitz MA: The metabolic activity of

Mycobacterium tuberculosis, assessed by use of a novel
inducible GFP expression system, correlates with its capacity
to inhibit phagosomal maturation and acidification in human
macrophages. Mol Microbiol 2008, 68:1047-1060.
Although the central message of this paper is not on phagosomal escape
of M. tuberculosis, the reported observation that after 5 days of infection
25% of the infecting M. tuberculosis cells were found without phagosomal membrane in the cytoplasm of human macrophages, confirms the
data on phagosomal escape proposed by van der Wel and colleagues.
49. Majlessi L, Brodin P, Brosch R, Rojas MJ, Khun H, Huerre M,
Cole ST, Leclerc C: Influence of ESAT-6 Secretion System 1
(RD1) of Mycobacterium tuberculosis on the interaction
between mycobacteria and the host immune system.
J Immunol 2005, 174:3570-3579.
50. Stanley SA, Johndrow JE, Manzanillo P, Cox JS: The Type I IFN
 response to infection with Mycobacterium tuberculosis
requires ESX-1-mediated secretion and contributes to
pathogenesis. J Immunol 2007, 178:3143-3152.
This paper suggests that the ESX-1 secretion system secretes effectors
into the cytoplasm of infected macrophages, thereby triggering the type I
IFN response for the manipulation of host immunity.

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