Biology subjects

Natarajan, G.

Publications and source records attributed to Natarajan, G..

5 recordsLinked to original sources

bioRxiv2020

Structural Basis of Complex Formation Between Mitochondrial Anion Channel VDAC1 and Hexokinase-II

Complex formation between hexokinase-II (HKII) and the mitochondrial channel VDAC1 plays a crucial role in regulating cell growth and survival; however, structural details of this complex remain elusive. We hypothesize that a conserved, hydrophobic helix (H-anchor) of HKII first inserts into the outer membrane of mitochondria (OMM) and then interacts with VDAC1 on the cytosolic leaflet of OMM to form a binary complex. To systematically investigate this process, we adopted a hybrid approach: 1) the membrane binding of HKII was first described with molecular dynamics (MD) simulations employing a membrane mimetic model with enhanced lipid diffusion, then 2) the resulting membrane-bound HKII was used to form complex with VDAC1 in millisecond-scale Brownian dynamics (BD) simulations. We show that H-anchor inserts its first 10 residues into the membrane, substantiating previous experimental findings. The insertion depth of the H-anchor was used to derive positional restraints in subsequent BD simulations to preserve the membrane-bound pose of HKII during the formation of the HKII/VDAC1 binary complex. Multiple BD-derived structural models were further refined with MD simulations, resulting in one stable complex. A major feature in the complex is the partial (not complete) blockade of VDAC1s permeation pathway by HKII, a result supported by our comparative electrophysiological measurements of the channel in the presence and absence of HKII. Additionally, we showed how VDAC1 phosphorylation disrupts HKII binding, a feature that is verified by our electrophysiology recordings and have implications in mitochondria-mediated cell death.

biophysics↗

bioRxiv2020

Species level enrichment of AnAOB and associated growth morphology under the effect of key metabolites

Nineteen anaerobic ammonium oxidizing bacteria (AnAOB) species have been identified, yet the environmental factors that select for each species within a specialized ecological niche have not been described. We enriched AnAOB from a single inoculum under standard enrichment conditions (reactor R1) with stepwise increase in nitrite and ammonia concentration, nitric oxide (NO) supplementation (reactor R2), or with complex organic carbon using wastewater collected from mainstream wastewater treatment facility (reactor R3). AnAOB were enriched up to 80%, 90% and 50% relative abundance in R1, R2 and R3 respectively. Candidatus Brocadia caroliniensis predominated in all reactors, but a shift towards Ca. Brocadia sinica was consistently observed with increasing ammonium and nitrite concentrations beyond 270 mg NH4-N L-1 and 340 mg NO2-N L-1, respectively. In the presence of NO, growth of heterotrophs were inhibited, and Ca. Jettenia could coexist with Ca. B. caroliniensis before diminishing when nitrite increased to 160 mg NO2-N L-1. In contrast, supplementation of organic carbon led to the emergence of heterotrophic communities that coevolved with Ca. B. caroliniensis. Ca. B. caroliniensis and Ca. Jettenia preferentially form biofilms on reactor surfaces, whereas Ca. Brocadia sinica forms granules in suspension. Our results thus indicate that multiple AnAOB species co-exist and occupy sub-niches in anaerobic ammonium oxidation reactors, that the dominant population can be reversibly shifted by, for example, changing the nitrogen load (i.e. high nitrite concentration favors Ca. Brocadia caroliniensis), and that speciation has implications for wastewater process design, with the optimum cell immobilization strategy (i.e. carriers vs granules) dependent on which species dominates. IMPORTANCEThis study demonstrates how to reversibly and predictably shift dominant anammox population using operating parameters (e.g. high nitrite concentration favours Ca. Brocadia sinica), and that species selection has implications for wastewater process design, illustrated here in terms of dependence of optimum cell immobilization strategy (i.e. carriers vs granules) on which species dominates. The research informs the characterization of AnAOBs at species level as well process design and control strategies targeting Anammox species population dynamics in full scale waste water treatment systems.

ecology↗

bioRxiv2018

Improving recovery of member genomes from enrichment reactor microbial communities using MinION--based long read metagenomics

New long read sequencing technologies offer huge potential for effective recovery of complete, closed genomes. While much progress has been made on cultured isolates, the ability of these methods to recover genomes of member taxa in complex microbial communities is less clear. Here we examine the ability of long read data to recover genomes from enrichment reactor metagenomes. Such modified communities offer a moderate level of complexity compared to the source communities and so are realistic, yet tractable, systems to use for this problem. We sampled an enrichment bioreactor designed to target anaerobic ammonium-oxidising bacteria (AnAOB) and sequenced genomic DNA using both short read (Illumina 301bp PE) and long read data (MinION Mk1B) from the same extraction aliquot. The community contained 23 members, of which 16 had genome bins defined from an assembly of the short read data. Two distinct AnAOB species from genus Candidatus Brocadia were present and had complete genomes, of which one was the most abundant member species in the community. We can recover a 4Mb genome, in 2 contigs, of long read assembled sequence that is unambiguously associated with the most abundant AnAOB member genome. We conclude that obtaining near closed, complete genomes of members of low-medium microbial communities using MinION long read sequence is feasible.

genomics↗

bioRxiv2018

Apparent oxygen half saturation constant for nitrifiers: genus specific, inherent physiological property, or artefact of colony morphology?

We report that a single Nitrospira sublineage I OTU performs nitrite oxidation in several full-scale domestic wastewater treatment plants (WWTPs) in the tropics (29-31 {degrees}C). Contrary to the prevailing theory for the relationship between nitrite oxidizing bacteria (NOB) and ammonia oxidizing bacteria (AOB), members of the Nitrospira sublineage I OTU had an apparent half saturation coefficient, Ks(app) lower than that of the full-scale domestic activated sludge cohabitant AOB (0.09 {+/-} 0.02 g O2 m-3 versus 0.3 {+/-} 0.03 g O2 m-3). Paradoxically, NOB may thus thrive under conditions of low oxygen supply. Low dissolved oxygen (DO) conditions could enrich for and high aeration inhibit the NOB in a long-term lab-scale reactor. The relative abundance of Nitrospira gradually decreased with increasing DO until it was washed out. Nitritation was sustained even after the DO was lowered subsequently. Based on 3D-fluorescence in situ hybridization (FISH) image analysis, the morphologies of AOB and NOB microcolonies responded to DO levels in accordance with their apparent oxygen half saturation constant Ks(app). When exposed to the same oxygenation level, NOB formed densely packed spherical clusters with a low surface area-to-volume ratio compared to the Nitrosomonas-like AOB clusters, which maintained a porous and non-spherical morphology. Microcolony morphology is thus a way for AOB and NOB to regulate oxygen exposure and sustain the mutualistic interaction. However, short-term high DO exposure can select for AOB and against NOB in full-scale domestic WWTPs and such population dynamics depend on which specific AOB and NOB species predominate under given environmental conditions.

systems biology↗

bioRxiv2017

Draft Genome Sequence Of A Candidatus Brocadia Bacterium Enriched From Tropical-Climate Activated Sludge

We present the draft genome of an anaerobic ammonium-oxidizing (anammox) bacterium, cluster III Candidatus Brocadia, which was enriched in an anammox reactor. A 3.2 Mb genome sequence comprising 168 contigs was assembled, in which 2,765 gene-coding regions, 47 tRNAs, and 5S, 16S and 23S ribosomal RNAs were annotated. No evidence for the presence of a nitric oxide-forming nitrite reductase was found.

microbiology↗