Expression and functional characterisation of TNC, a high-affinity nickel transporter from Neurospora crassa

Tiwari, Anand; Korripally, Premsagar; Adhikarla, Haritha; Patnala, Kiranmayi; Pamarthi, Maruthi Mohan; Bhanoori, Manjula

doi:10.1016/j.fgb.2011.07.006

Cited by 8 publications

(1 citation statement)

References 26 publications

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“…Therefore, the presence of a low-capacity nickel uptake system may answer the physiological requirements of the organism. Several studies have shown that expression of NiCoTs from different organisms in E. coli strains can improve bioaccumulation of nickel, which can be a potential application for the purpose of bioremediation of nickel-polluted environments [39,45,46]. In the study by Zhang et al, Ni 2+ uptake increased from 3.7 mg g −1 of host strain to 11 .3 mg g −1 when a NiCoT from S. aureus was expressed in E. coli BL21 [47].…”

Section: Discussionmentioning

confidence: 99%

A NiCoT family metal transporter of Mycobacterium tuberculosis (Rv2856/NicT) behaves as a drug efflux pump that facilitates cross-resistance to antibiotics

et al. 2022

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Metals often act as a facilitator in the proliferation and persistence of antibiotic resistance. Efflux pumps play key roles in the co-selection of metal and antibiotic resistance. Here, we report the ability of a putative nickel/cobalt transporter (NiCoT family), Rv2856 or NicT of Mycobacterium tuberculosis (Mtb), to transport metal and antibiotics and identified some key amino acid residues that are important for its function. Ectopic expression of NicT in Escherichia coli CS109 resulted in the increase of intracellular nickel uptake. Additionally, enhanced tolerance towards several antibiotics (norfloxacin, sparfloxacin, ofloxacin, gentamicin, nalidixic acid and isoniazid) was observed with NicT overexpression in E. coli and Mycobacterium smegmatis . A comparatively lower intracellular accumulation of norfloxacin upon NicT overexpression than that of the cells without NicT indicated the involvement of NicT in an active efflux process. Although expression of NicT did not alter the sensitivity towards kanamycin, doxycycline, tetracycline, apramycin, neomycin and ethambutol, the presence of a sub-inhibitory dose of Ni2+ resulted in the manifestation of low-level tolerance towards these drugs. Further, substitution of four residues (H77I, D82I, H83L and D227I) in the conserved regions of NicT by isoleucine and leucine resulted in reduced to nearly complete loss of the transport function for both metals and antimicrobials. Therefore, the study suggests that nickel transporter Rv2856/NicT may actively export different drugs and the presence of nickel might drive the cross-resistance to some of the antibiotics.

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Section: Discussionmentioning

confidence: 99%

A NiCoT family metal transporter of Mycobacterium tuberculosis (Rv2856/NicT) behaves as a drug efflux pump that facilitates cross-resistance to antibiotics

et al. 2022

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Nickel, Cobalt Transport in Prokaryotes

Eitinger

2004

Encyclopedia of Inorganic and Bioinorganic Chemistry

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Nickel and cobalt ions are required by prokaryotes for incorporation into diverse enzymes involved in central metabolic reactions. The metal ions are taken up from the environment by selective high‐affinity primary and secondary active transport systems. These systems discriminate between Ni 2+ and Co 2+ on the one side and other divalent transition metal ions on the other side. Despite similar properties of Ni 2+ and Co 2+ , a couple of primary and secondary uptake systems exist that efficiently distinguish even between those two cations. Relevant transporters include canonical and energy‐coupling factor‐type ATP‐binding cassette importers and members of the nickel/cobalt transporter family and relatives of the latter. Recent advances include the discovery of active transport of nickel and cobalt across the outer membrane of gram‐negative bacteria by TonB‐dependent systems and the tentative identification of an unusual metallophore for nickel uptake. Export systems that avoid cellular overload with nickel and cobalt ions comprise multimeric systems of the resistance‐nodulation‐cell division family, secondary active systems from different subgroups, and P‐type ATPases.

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The NcZrg-17 gene of Neurospora crassa encodes a cation diffusion facilitator transporter required for vegetative development, tolerance to endoplasmic reticulum stress and cellulose degradation under low zinc conditions

2017

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The Neurospora crassa gene NcZrg-17 encodes a membrane protein with homology to the cation diffusion facilitator (CDF) family of transporters. We analyzed the phenotypic and functional characteristics of ΔNcZrg-17 and the implications of these characteristics in vivo. The ΔNcZrg-17 mutant showed several phenotypes that are zinc suppressible such as reduced growth rate, short aerial hyphae, increased hyphal branching, early and enhanced conidiation and delayed conidial germination. Furthermore, the NcZrg-17 gene was found to be crucial for survival in the presence of endoplasmic reticulum (ER) stress inducing chemical agents. In addition, we found that ΔNcZrg-17 mutant is defective in protein secretion on cellulose media under low zinc conditions, pointing towards a physiological role for NcZrg-17 in N. crassa. A gradual and delayed transcriptional upregulation (~ threefold) of NcZrg-17 on exposure to low zinc suggests its role in adaptation to low zinc rather than zinc homeostasis. Together our findings support a function of NcZrg-17 in normal vegetative growth, tolerance to ER stress and degradation of cellulose under low zinc conditions in N. crassa.

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Expression and functional characterisation of TNC, a high-affinity nickel transporter from Neurospora crassa

Cited by 8 publications

References 26 publications

A NiCoT family metal transporter of Mycobacterium tuberculosis (Rv2856/NicT) behaves as a drug efflux pump that facilitates cross-resistance to antibiotics

A NiCoT family metal transporter of Mycobacterium tuberculosis (Rv2856/NicT) behaves as a drug efflux pump that facilitates cross-resistance to antibiotics

Nickel, Cobalt Transport in Prokaryotes

The NcZrg-17 gene of Neurospora crassa encodes a cation diffusion facilitator transporter required for vegetative development, tolerance to endoplasmic reticulum stress and cellulose degradation under low zinc conditions

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