BACTERIOFERRITIN
BACTERIOFERRITIN
BACTERIOFERRITIN
Department of Biochemistry, National JALMA Institute for Leprosy and Other Mycobacterial Diseases, Agra, India
Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 1 June 2017 | Volume 7 | Article 240
Sharma and Bisht Iron Storage Proteins vs. Aminoglycosides Resistance
they typically inhibit protein synthesis by interacting with protein concentration of iron. They depicted that increasing iron
translational machinery. Two dimensional gel electrophoresis concentration (absence of bacterioferritin and ferritin) decreased
coupled with mass spectrometry is the best accepted approach resistance to various anti-TB drugs including aminoglycosides
for expression proteomics (Lata et al., 2015a,b; Sharma et al., as well as fluoroquinolones (Pandey and Rodriguez, 2012)
2015b, 2016b; Sharma and Bisht, 2016). Since last decade and suggested that ferritin and bacterioferritin are not only
a panel of proteomics and bioinformatics studies related to mandatory to maintain iron homeostasis but also make
aminoglycosides resistance have been accumulated (Sharma M. tuberculosis resistant to aminoglycosides. Kurthkoti et al
et al., 2010, 2014, 2015b,a, 2016a,b; Kumar et al., 2013; suggested that iron-dependent regulator IdeR induces ferritin
Sharma and Bisht, 2017a,b). Here we emphasized on the M. (bfrB) expression by alleviating Lsr2 repression in M. tuberculosis
tuberculosis iron storage proteins (bacterioferritin and ferritin) (Kurthkoti et al., 2015). WhiB7 (Rv3197A) is Fe-S cluster-bound
and their interactive protein partners which might be involved protein (transcriptional regulatory protein), which not only
in pathogenesis, virulence and drug resistance. associates with aminoglycosides resistance but also predicted to
Iron is an essential entity for metabolism of the biological cells be in the IdeR|Rv2711 regulon (Gold et al., 2001; Morris et al.,
and hence crucial for the chemistry of life. Iron assimilation, 2005). Recently Kuberl et al. (2016) suggested that pupylation
storage and their utilization play a crucial role not only machinery maintains the iron homeostasis by targeting iron
in pathogenesis/pathobiology (growth, survival, virulence and storage proteins (Kuberl et al., 2016).
latency) but also in emergence of aminoglycosides drug In our previous studies of expression proteomics we
resistance strains of M. tuberculosis (Reddy et al., 2012; Kumar have discovered, that bacterioferritin (Rv1876) and ferritin
et al., 2013; Sharma et al., 2015b). Recently, Khare et al. (Rv3841) were overexpressed in aminoglycosides (amikacin and
(2017) reported that iron storage proteins are involved in kanamycin) resistant M. tuberculosis clinical isolates (Kumar
maintaining iron homeostasis in M. tuberculosis (Khare et al., et al., 2013; Sharma et al., 2015b). Molecular docking revealed
2017). Bacterioferritin (Rv1876) and ferritin (Rv3841) are unique that aminoglycosides drugs (AK and KM) bind to conserved
to maintain iron storage as well as homeostasis in M. tuberculosis. bacterioferritin domain of Rv1876 and ferritin domain of Rv3841
In mycobacteria, various genes products and their interactive and suggested that overexpression of these proteins might be
partners required for high affinity iron acquisition have been to neutralize/modulate the drug effect and could be involved
identified such as siderophore production, uptake of ferric- in aminoglycosides resistance mechanisms of M. tuberculosis
siderophores, production of iron storage proteins and uptake (Kumar et al., 2013; Sharma et al., 2015b). Although the enzymes
of heme. Production, storage and function of iron uptake and their connected pathways involved in iron metabolism
mechanisms are controlled by a regulatory protein IdeR (Gold in M. tuberculosis are well recognized, still our information
et al., 2001). Heme is the preferable iron source for M. related to iron transportation, trafficking, iron dependent
tuberculosis and its acquisition is done by a biosynthetic enzyme post-transcriptional & translational regulations and protein-
ferrochelatase (Rv1485). Rv1485 catalyzes the last step of heme protein interactions in mycobacteria is inadequate. Recently
biosynthesis in which iron is interleaved to protoporfhyrin IX Sharma et al. (2016a) reported that inducible overexpression
to form protoheame (Dailey and Dailey, 2002). It is essential of recombinant ferritin in E. coli (BL21) leads to shift in MIC
because it supplies the heme which is a preferred iron source of AK & KM and suggested their probable roles in conferring
for M. tuberculosis (Parish et al., 2005) and serves as a cofactor aminoglycosides resistance (Sharma et al., 2016a). Consequently,
for various metabolic enzymes such as catalase-peroxidase and proteins involved in iron storage, assimilation, regulation, uptake
DosS/DosT two component system [active sites contains heme] and their utilization can be a promising anti-mycobacterial
(Svistunenko, 2005). targets against the drug resistant tuberculosis.
In pathogenic mycobacteria, usually high affinity systems
are essential for maintenance of an infection, virulence BACTERIOFERRITIN AND FERRITIN
and resistance. Greater definition of the functions of both
the identified genes and their products, ferritin (bfrB) and
PROTEIN-PROTEIN INTERACTION:
bacterioferritin (bfrA) will refine our understanding of UNLOCK THE SECRETS OF IRON
mycobacterial iron acquisition and the interplay between RELATED PATHWAYS IN
components of the iron systems have increased intensities AMINOGLYCOSIDES DRUG RESISTANCE
under iron-rich and decreased intensities under iron deprived
conditions (Pandey and Rodriguez, 2012). Under iron-rich STRING-10 is an online server which is used to predict the
conditions, M. tuberculosis represses iron acquisition and interacting partners of the iron storage proteins [bacterioferritin
induces iron storage proteins suggesting the significant role (Rv1876) and ferritin (Rv3841)] (Sharma et al., 2016a,b;
of iron storage proteins in iron homeostasis. M. tuberculosis Sharma and Bisht, 2017a). “STRING uses a combination
usually synthesizes two iron storage proteins: ferritin (bfrB) of prediction approaches and an integration of other
and bacterioferritin (bfrA). BfrB is mandatory to overcome information (neighborhood, transferred neighborhood,
iron limitation and defense against oxidative stress, whereas gene fusion, co-occurrence, co-expression, experiments,
bfrA is superfluous for victorious adaptation to those stresses. databases, text mining). Network was made at medium
Studies of M. tuberculosis lacking bfrB gene (which encode the confidence level (0.400) allowing all active prediction methods,
ferritin an iron storage protein) reported increased intracellular which corresponds to approximately 50% possibility of
Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 2 June 2017 | Volume 7 | Article 240
Sharma and Bisht Iron Storage Proteins vs. Aminoglycosides Resistance
FIGURE 1 | (A) STRING analysis reveals the interaction partners of M. tuberculosis H37Rv bacterioferritin and ferritin protein. (B) The score for each interaction
partner is assigned and shown in the table inbuilt in figure. The highest score for hemH association was found to be 0.816 followed by Rv1877, a probable transporter
(its confidence score of association was 0.616). The confidence score for association for tig corresponds to 0.577, and for molecular cheprone (groEL1 & hsp65) was
found to be 0.469.
Frontiers in Cellular and Infection Microbiology | www.frontiersin.org 3 June 2017 | Volume 7 | Article 240
Sharma and Bisht Iron Storage Proteins vs. Aminoglycosides Resistance
hypotheticals and regulatory proteins. Hypothetical protein (Sharma et al., 2015b) and suggested that trigger factor
(Rv1877), a Probable conserved integral membrane protein, might trigger expression of bacterioferritin and ferritin
ferrochelatase and trigger factor might be potential novel and their interactive partners which could be involved in
targets predicted by bacterioferritin and ferritin protein- aminoglycosides drug resistance. We suggested that cumulative
protein interaction. Rv1877 having 14-transmembrane helixes effect of iron storage proteins (bacterioferritin and ferritin)
(TMH), possibly involved in transport of drug across the and their interactive partners {hypothetical protein (Rv1877),
membrane and could be a potential drug target against the ferrochelatase, trigger factor and others} might be involved in
aminoglycosides drug resistance. Rv1877 is the homologous various stress, and aminoglycosides drug resistance. Predicted
of lfrA and deletion of this gene increased the susceptibility pupylation sites in bacterioferritin and ferritin also suggested
against various antibiotics including the aminoglycosides in its involvement not only in iron homeostasis but also in
mycobacteria (Li et al., 2004). Recently, Mehra et al. (2016) aminoglycosides resistance (Kuberl et al., 2016; Sharma et al.,
reported that Rv1877 is the part of ABC transporter and 2016a). Further detailed and in-depth investigations of these
predicted its involvement in drug resistance (Mehra et al., interactome and pupylome could explore the aminoglycosides
2016). Ferrochelatase involved in heme biosynthetic pathways resistance and might be used as potential drug targets against
is a preferred iron source for M. tuberculosis. Trigger factor this issue.
which not only having chaperone activity but also involved
in eliciting the protein expression and their export which AUTHOR CONTRIBUTIONS
could maintain the 3D conformation of newly synthesized
proteins and prevents misfolding as well as promotes the DS design the concept and wrote the manuscript. DS and DB
refolding of unfolded polypeptides generated under stressed finalized the manuscript.
conditions. Bhuwan et al. (2016) reported and validated the
STRING in the interaction of RipA with Chaperone MoxR1 ACKNOWLEDGMENTS
(Bhuwan et al., 2016). Sharma et al. (2015b) reported that
trigger factor, bacterioferritin and ferritin were overexpressed The authors are grateful to Director, NJIL, & OMD for the
in aminoglycosides resistant M. tuberculosis clinical isolates support. DS is ICMR-PDFs (ICMR, New Delhi).
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Sharma, D., Lata, M., Singh, R., Deo, N., Venkatesan, K., and Bisht, D. (2016b). Conflict of Interest Statement: The authors declare that the research was
Cytosolic proteome profiling of aminoglycosides resistant Mycobacterium conducted in the absence of any commercial or financial relationships that could
tuberculosis clinical isolates using MALDI-TOF/MS. Front. Microbiol. 7:1816. be construed as a potential conflict of interest.
doi: 10.3389/fmicb.2016.01816
Sharma, D., Shankar, H., Lata, M., Joshi, B., Venkatesan, K., and Bisht, Copyright © 2017 Sharma and Bisht. This is an open-access article distributed
D. (2014). Culture filtrate proteome analysis of aminoglycoside resistant under the terms of the Creative Commons Attribution License (CC BY). The use,
clinical isolates of Mycobacterium tuberculosis. BMC Infect. Dis. 14:P60. distribution or reproduction in other forums is permitted, provided the original
doi: 10.1186/1471-2334-14-S3-P60 author(s) or licensor are credited and that the original publication in this journal
Sharma, P., Kumar, B., Gupta, Y., Singhal, N., Katoch, V. M., Venkatesan, is cited, in accordance with accepted academic practice. No use, distribution or
K., et al. (2010). Proteomic analysis of streptomycin resistant and reproduction is permitted which does not comply with these terms.
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