Satellite remote sensing reveals a positive impact of living oyster reefs on microalgal biofilm development

Satellite remote sensing (RS) is routinely used for the large-scale monitoring of microphytobenthos (MPB) biomass in intertidal mudflats and has greatly improved our knowledge of MPB spatio-temporal variability and its potential drivers. Processes operating on smaller scales however, such as the impact of benthic macrofauna on MPB development, to date remain underinvestigated. In this study, we analysed the influence of wild Crassostrea gigas oyster reefs on MPB biofilm development using multispectral RS. A 30-year time series (1985-2015) combining high-resolution (30 m) Landsat and SPOT data was built in order to explore the relationship between C. gigas reefs and MPB spatial distribution and seasonal dynamics, using the normalized difference vegetation index (NDVI). Emphasis was placed on the analysis of a before-after control-impact (BACI) experiment designed to assess the effect of oyster killing on the surrounding MPB biofilms. Our RS data reveal that the presence of oyster reefs positively affects MPB biofilm development. Analysis of the historical time series first showed the presence of persistent, highly concentrated MPB patches around oyster reefs. This observation was supported by the BACI experiment which showed that killing the oysters (while leaving the physical reef structure, i.e. oyster shells, intact) negatively affected both MPB biofilm biomass and spatial stability around the reef. As such, our results are consistent with the hypothesis of nutrient input as an explanation for the MPB growth-promoting effect of oysters, whereby organic and inorganic matter released through oyster excretion and biodeposition stimulates MPB biomass accumulation. MPB also showed marked seasonal variations in biomass and patch shape, size and degree of aggregation around the oyster reefs. Seasonal variations in biomass, with higher NDVI during spring and autumn, were consistent with those observed on broader scales in other European mudflats. Our study provides the first multi-sensor RS satellite evidence of the promoting and structuring effect of oyster reefs on MPB biofilms

Satellite remote sensing reveals a positive impact of living oyster reefs on microalgal biofilm development

Satellite remote sensing (RS) is routinely used for the large-scale monitoring of microphytobenthos (MPB) biomass in intertidal mudflats and has greatly improved our knowledge of MPB spatio-temporal variability and its potential drivers. Processes operating on smaller scales however, such as the impact of benthic macrofauna on MPB development, to date remain underinvestigated. In this study, we analysed the influence of wild Crassostrea gigas oyster reefs on MPB biofilm development using multispectral RS. A 30-year time series (1985–2015) combining high-resolution (30 m) Landsat and SPOT data was built in order to explore the relationship between C. gigas reefs and MPB spatial distribution and seasonal dynamics, using the normalized difference vegetation index (NDVI). Emphasis was placed on the analysis of a before–after control-impact (BACI) experiment designed to assess the effect of oyster killing on the surrounding MPB biofilms. Our RS data reveal that the presence of oyster reefs positively affects MPB biofilm development. Analysis of the historical time series first showed the presence of persistent, highly concentrated MPB patches around oyster reefs. This observation was supported by the BACI experiment which showed that killing the oysters (while leaving the physical reef structure, i.e. oyster shells, intact) negatively affected both MPB biofilm biomass and spatial stability around the reef. As such, our results are consistent with the hypothesis of nutrient input as an explanation for the MPB growth-promoting effect of oysters, whereby organic and inorganic matter released through oyster excretion and biodeposition stimulates MPB biomass accumulation. MPB also showed marked seasonal variations in biomass and patch shape, size and degree of aggregation around the oyster reefs. Seasonal variations in biomass, with higher NDVI during spring and autumn, were consistent with those observed on broader scales in other European mudflats. Our study provides the first multi-sensor RS satellite evidence of the promoting and structuring effect of oyster reefs on MPB biofilms

Antagonistic Changes in Sensitivity to Antifungal Drugs by Mutations of an Important ABC Transporter Gene in a Fungal Pathogen

Fungal pathogens can be lethal, especially among immunocompromised populations, such as patients with AIDS and recipients of tissue transplantation or chemotherapy. Prolonged usage of antifungal reagents can lead to drug resistance and treatment failure. Understanding mechanisms that underlie drug resistance by pathogenic microorganisms is thus vital for dealing with this emerging issue. In this study, we show that dramatic sequence changes in PDR5, an ABC (ATP-binding cassette) efflux transporter protein gene in an opportunistic fungal pathogen, caused the organism to become hypersensitive to azole, a widely used antifungal drug. Surprisingly, the same mutations conferred growth advantages to the organism on polyenes, which are also commonly used antimycotics. Our results indicate that Pdr5p might be important for ergosterol homeostasis. The observed remarkable sequence divergence in the PDR5 gene in yeast strain YJM789 may represent an interesting case of adaptive loss of gene function with significant clinical implications

The ATP-Binding Cassette Proteins of the Deep-Branching Protozoan Parasite Trichomonas vaginalis

The ATP binding cassette (ABC) proteins are a family of membrane transporters and regulatory proteins responsible for diverse and critical cellular process in all organisms. To date, there has been no attempt to investigate this class of proteins in the infectious parasite Trichomonas vaginalis. We have utilized a combination of bioinformatics, gene sequence analysis, gene expression and confocal microscopy to investigate the ABC proteins of T. vaginalis. We demonstrate that, uniquely among eukaryotes, T. vaginalis possesses no intact full-length ABC transporters and has undergone a dramatic expansion of some ABC protein sub-families. Furthermore, we provide preliminary evidence that T. vaginalis is able to read through in-frame stop codons to express ABC transporter components from gene pairs in a head-to-tail orientation. Finally, with confocal microscopy we demonstrate the expression and endoplasmic reticulum localization of a number of T. vaginalis ABC transporters

The λ Red Proteins Promote Efficient Recombination between Diverged Sequences: Implications for Bacteriophage Genome Mosaicism

Genome mosaicism in temperate bacterial viruses (bacteriophages) is so great that it obscures their phylogeny at the genome level. However, the precise molecular processes underlying this mosaicism are unknown. Illegitimate recombination has been proposed, but homeologous recombination could also be at play. To test this, we have measured the efficiency of homeologous recombination between diverged oxa gene pairs inserted into λ. High yields of recombinants between 22% diverged genes have been obtained when the virus Red Gam pathway was active, and 100 fold less when the host Escherichia coli RecABCD pathway was active. The recombination editing proteins, MutS and UvrD, showed only marginal effects on λ recombination. Thus, escape from host editing contributes to the high proficiency of virus recombination. Moreover, our bioinformatics study suggests that homeologous recombination between similar lambdoid viruses has created part of their mosaicism. We therefore propose that the remarkable propensity of the λ-encoded Red and Gam proteins to recombine diverged DNA is effectively contributing to mosaicism, and more generally, that a correlation may exist between virus genome mosaicism and the presence of Red/Gam-like systems

Rational Mutational Analysis of a Multidrug MFS Transporter CaMdr1p of Candida albicans by Employing a Membrane Environment Based Computational Approach

CaMdr1p is a multidrug MFS transporter of pathogenic Candida albicans. An over-expression of the gene encoding this protein is linked to clinically encountered azole resistance. In-depth knowledge of the structure and function of CaMdr1p is necessary for an effective design of modulators or inhibitors of this efflux transporter. Towards this goal, in this study, we have employed a membrane environment based computational approach to predict the functionally critical residues of CaMdr1p. For this, information theoretic scores which are variants of Relative Entropy (Modified Relative Entropy REM) were calculated from Multiple Sequence Alignment (MSA) by separately considering distinct physico-chemical properties of transmembrane (TM) and inter-TM regions. The residues of CaMdr1p with high REM which were predicted to be significantly important were subjected to site-directed mutational analysis. Interestingly, heterologous host Saccharomyces cerevisiae, over-expressing these mutant variants of CaMdr1p wherein these high REM residues were replaced by either alanine or leucine, demonstrated increased susceptibility to tested drugs. The hypersensitivity to drugs was supported by abrogated substrate efflux mediated by mutant variant proteins and was not attributed to their poor expression or surface localization. Additionally, by employing a distance plot from a 3D deduced model of CaMdr1p, we could also predict the role of these functionally critical residues in maintaining apparent inter-helical interactions to provide the desired fold for the proper functioning of CaMdr1p. Residues predicted to be critical for function across the family were also found to be vital from other previously published studies, implying its wider application to other membrane protein families

Alternative-NHEJ Is a Mechanistically Distinct Pathway of Mammalian Chromosome Break Repair

Characterizing the functional overlap and mutagenic potential of different pathways of chromosomal double-strand break (DSB) repair is important to understand how mutations arise during cancer development and treatment. To this end, we have compared the role of individual factors in three different pathways of mammalian DSB repair: alternative-nonhomologous end joining (alt-NHEJ), single-strand annealing (SSA), and homology directed repair (HDR/GC). Considering early steps of repair, we found that the DSB end-processing factors KU and CtIP affect all three pathways similarly, in that repair is suppressed by KU and promoted by CtIP. In contrast, both KU and CtIP appear dispensable for the absolute level of total-NHEJ between two tandem I-SceI–induced DSBs. During later steps of repair, we find that while the annealing and processing factors RAD52 and ERCC1 are important to promote SSA, both HDR/GC and alt-NHEJ are significantly less dependent upon these factors. As well, while disruption of RAD51 causes a decrease in HDR/GC and an increase in SSA, inhibition of this factor did not affect alt-NHEJ. These results suggest that the regulation of DSB end-processing via KU/CtIP is a common step during alt-NHEJ, SSA, and HDR/GC. However, at later steps of repair, alt-NHEJ is a mechanistically distinct pathway of DSB repair, and thus may play a unique role in mutagenesis during cancer development and therapy

ABC Transporter Pdr10 Regulates the Membrane Microenvironment of Pdr12 in Saccharomyces cerevisiae

The eukaryotic plasma membrane exhibits both asymmetric distribution of lipids between the inner and the outer leaflet and lateral segregation of membrane components within the plane of the bilayer. In budding yeast (Saccharomyces cerevisiae), maintenance of leaflet asymmetry requires P-type ATPases, which are proposed to act as inward-directed lipid translocases (Dnf1, Dnf2, and the associated protein Lem3), and ATP-binding cassette (ABC) transporters, which are proposed to act as outward-directed lipid translocases (Pdr5 and Yor1). The S. cerevisiae genome encodes two other Pdr5-related ABC transporters: Pdr10 (67% identity) and Pdr15 (75% identity). We report the first analysis of Pdr10 localization and function. A Pdr10-GFP chimera was located in discrete puncta in the plasma membrane and was found in the detergent-resistant membrane fraction. Compared to control cells, a pdr10∆ mutant was resistant to sorbate but hypersensitive to the chitin-binding agent Calcofluor White. Calcofluor sensitivity was attributable to a partial defect in endocytosis of the chitin synthase Chs3, while sorbate resistance was attributable to accumulation of a higher than normal level of the sorbate exporter Pdr12. Epistasis analysis indicated that Pdr10 function requires Pdr5, Pdr12, Lem3, and mature sphingolipids. Strikingly, Pdr12 was shifted to the detergent-resistant membrane fraction in pdr10∆ cells. Pdr10 therefore acts as a negative regulator for incorporation of Pdr12 into detergent-resistant membranes, a novel role for members of the ABC transporter superfamily