coli lac promoter. The lipA gene was removed from the resulting plasmid

pBBLCAH by BamHI digestion and recircularization, yielding pBBLCH, which harbours a synthetic lipC/lipH operon under the transcriptional control of Plac and PT7. M9-minimal broth medium contained, per litre, 4 g glucose; 0.25 g MgSO4; 0.02 g CaCl2; 7 g Na2HPO4; 0.3 g KH2PO4; 0.5 g NaCl; 1 g NH4Cl; and 0.3% w/v agar (Oxoid). Swim plates were inoculated from overnight cultures of bacteria grown on LB plates (1.5% w/v agar) at 37 °C with a sterile toothpick. Plates were then incubated at 37 °C for 24 h. The M9-minimal broth medium described above was used with NH4Cl replaced by 0.05% w/v glutamate and solidified by the addition of 0.5% w/v agar (Oxoid). Plates were briefly dried before use, inoculated and incubated at 37 °C for 36 h. LB medium contained, per litre, 10 g see more tryptone; 5 g yeast extract; 10 g NaCl; and 1.5% w/v agar (Oxoid). Plates were inoculated with a sharp toothpick stabbed

to the bottom of a Petri dish. The zone of motility at the agar/Petri dish interface was measured after incubation for 48 h at 37 °C. Bacteria were grown on swarming agar plates as described above and cells were removed from the edge of growing colonies. For thin sections, bacteria were prefixed in 3% glutaraldehyde Akt inhibitor in vivo in cacodylate buffer (200 mM), followed by osmium tetroxide 1% in cacodylate buffer and uranylacetate 3% in water. Samples were dehydrated in acetone and embedded in glycidether (EPon 812). Sections were stained with leadcitrate 4% and examined electromicroscopically with an EM 10 (Zeiss, Germany) at an acceleration voltage of 80 kV. For negative stain preparations, bacteria were mounted on copper grids and incubated for 40 s in uranylacetate 3%. Pseudomonas aeruginosa strains used in biofilm experiments were fluorescently tagged with gfp at an intergenic neutral chromosomal locus in a miniTn7 construct. Biofilms were grown in a modified NB medium [80 mg L−1

Lab Lemco broth (Oxoid); 1 mM MgCl2; 0.1 mM CaCl2; 50 mM NaCl; 6 g L−1 Na2HPO4·2H2O; 3 g L−1 KH2PO4; and nonchelated trace metals] on glass cover slips in aminophylline flow chambers at 30 °C as described previously (Klausen et al., 2003). The flow chambers were inoculated by injecting 350 μL of an overnight culture diluted to an OD600 nm of 0.001 into individual flow channels with dimensions of 1 × 4 × 40 mm. After inoculation, flow channels were left without flow for 1 h. Then, medium flow was started using a Watson Marlow 205S peristaltic pump to produce a flow rate of 3 mL h−1. Biofilm development was documented with an LSM 510 confocal laser scanning microscope (Zeiss) using a × 63/1.4 objective. Simulated three-dimensional images and sections were generated using the imaris software package (Bitplane). The biomass, surface coverage and roughness of two independent biofilms per strain were analysed using the comstat software.

albicans, which is responsible for at least 85% of human candidiasis (Rein, 1997), and A. neuii, which is the second most frequent microorganism isolated in the Ison and Hay grade II and III vaginal microbiota represented by bacterial vaginosis-related organisms (Verhelst et al., 2005) and has been also associated with bacterial vaginosis in women with intrauterine devices (Chatwani & Amin-Hanjani, 1994). Four of the lactobacilli enhanced the adherence of C. albicans and A. neuii to HeLa cells, which contrasts with previous findings, where pathogen adhesion inhibition was reported (Boris et al.,

1998; Osset et al., 2001). This fact suggests that this trait is strain specific. In fact, although the formation of a ternary complex pathogen–Lactobacillus–epithelial cell might enhance the antimicrobial effect of the lactic acid generated www.selleckchem.com/products/MK-2206.html by this GDC-0941 ic50 bacteria (Boris et al., 1997; Coudeyras et al., 2008), these ternary complexes could also enhance the pathogen adhesion as has been observed with Lactobacillus acidophilus and the adhesion of C. albicans to

the contraceptive vaginal ring (Chassot et al., 2010). Adhesion of A. neuii was very responsive to the addition of the extracellular proteins of the lactobacilli in a strain-dependent fashion. Five of them enhanced adsorption of the pathogen, thus reproducing the results obtained when whole bacterial cells were used. It is worth mentioning the extraordinary adhesion increment brought about by L. gasseri Lv19, which could be due to the secretion of an aggregation-promoting factor–like protein. In fact, it has

already been described that these factors act as bridges between pathogen and human cells (Marcotte et al., 2004). This synergistic effect has also been described for some exopolysaccharides produced by several probiotic Farnesyltransferase bacteria, including L. rhamnosus GG (Ruas-Madiedo et al., 2006). Interestingly, the extracellular proteins of L. plantarum Li69 and of L. salivarius Lv72 markedly inhibited the adhesion of A. neuii to HeLa cells. Among the different proteins secreted by these strains, several contained LysM domains, such as two peptidoglycan-binding proteins of Lv72. The LysM domain has been proposed to be the attachment site of the autolysin AcmA of Lactococcus lactis to peptidoglycan (Steen et al., 2003). Recently, an extracellular chitin-binding protein from L. plantarum, containing this domain, has been shown to attach to the cell surface and to selective bind N-acetylglucosamine-containing polymers (Sánchez et al., 2010). Notably, the Lv19 extracellular proteome, which enhanced A. neuii adhesion, did not include any LysM-bearing polypeptides. It is thus conceivable that binding of the LysM-bearing proteins to the A. neuii surface might block the ligands that recognize the surface of the HeLa cells, as already shown for other proteins (Spurbeck & Arvidson, 2010).

Although the CG and MDRD equations are widely used in HIV infection, they were derived from HIV-negative persons with

acute illnesses or ongoing CKD. The CG formula includes weight but not race, although it is known, for example, that African Americans have a higher muscle mass than Caucasians, while the MDRD and CKD-EPI formulae do not include weight, which may make it the equation of choice in observational studies if this variable is not measured for all patients. The CKD-EPI equation was derived in a less selected, but HIV-negative, population, Adriamycin in vivo and was found to be more accurate than existing estimates in the normal range [6]. None of the equations has yet been validated in HIV-positive persons. Comparison of the different equations has been

based on small numbers to date and results have been contradictory. In one small study, eGFR values derived using CG and MDRD were highly correlated [9], in another, MDRD was found to have the greatest accuracy [10], while another suggested CG was the best estimate of GFR in HIV-infected patients, as they are typically younger [11]. A comparison of the CG and MDRD equations in African patients enrolled in the Development of Antiretroviral Therapy trial suggested that the prevalence of moderate CKD was higher using MDRD compared with CG [12]. The difficulties do not end with having decided which method to use for calculating eGFR. In HIV-infected persons, the HIV Medicine Association of the Infectious Diseases Society of America has proposed five Palbociclib manufacturer SPTLC1 stages of CKD (see Table 1) [13] based on the National Kidney Foundation Kidney Disease Outcomes

Quality Initiative [14]. CKD is defined as either kidney damage (pathologic abnormalities or markers of damage) or GFR<60 mL/min/1.73 m2 for >3 months. An eGFR>60 mL/min/1.73 m2 is generally considered too inaccurate for routine clinical use and has been discouraged [15] with a recommendation that stages 1 and 2 are not used, in part because eGFR is particularly imprecise at low serum creatinine levels (normal to high GFR) [16]. In persons not infected with HIV, the staging system probably overestimates the prevalence of CKD and potentially misclassifies persons as having CKD in the absence of clinically relevant kidney disease [15]. There is little understanding of or research into outcomes following CKD in HIV-infected patients, or the prevalence of advanced (stage IV or V) CKD. Preliminary data from the EuroSIDA study suggested that up to one-third of patients resolved CKD [defined as either confirmed (≥3 months apart) eGFR≤60 mL/min/1.73 m2 for patients with baseline eGFR>60 mL/min/1.73 m2 or confirmed 25% decline in eGFR for patients with baseline eGFR≤60 mL/min/1.

Some residents, taking antipsychotics, were referred to a Psychiatry of Old Age Services (POAS) JQ1 consultant and their team who undertook a detailed review of antipsychotics with an aim to reduce inappropriate use. Following the review/MDT, options about which medicines should be stopped, changed or started were discussed with the resident and/or the family (in cases where the resident had no capacity to make informed decisions). The following questions were asked and discussed: Is the medication still needed i.e. currently treating or preventing disease? Does

the medicine still have benefits taking into consideration co-morbidities (e.g. palliative care)? Are there any medications not prescribed that the patient should be taking? Following any changes, residents were followed up monthly and post-review events were documented (i.e. any adverse event that was attributed to actions taken at the review). This abstract presents results from the first three (of twelve) care homes reviewed as part this project. Savings calculations were for medicines stopped/started and were based on the average savings from the pilot study (£32 per resident per month).2 Interim data: 86 residents have been reviewed over 16 sessions. They

were taking 749 medicines at the beginning of the review (8.7 medicines per resident). In total, 385 interventions were made including 241 medicines being stopped and 19 medicines started. At the end of Ganetespib solubility dmso the review, residents were taking 527 medicines (6.1 medicines per resident), resulting in a net reduction of 2.6 medicines per resident. There were 15 referrals to the POAS service. Etomidate Follow up for 44 residents has been undertaken and there have been 6 minor adverse events reported (e.g. rash following stopping antihistamine). Estimated monthly savings for 86 patients was £2,752, from medicines stopped/started. Other costs (pharmacist/GP/consultant time, hospital admissions) have yet to be determined, but will be

taken into consideration in an overall evaluation of the project. Through these reviews, residents were only prescribed medicines that were beneficial, appropriate and evidence based, ensuring full participation of the resident/family in any decisions made, with medicines deemed inappropriate or unnecessary being discontinued. Follow-up identified few minor events from discontinuing over two hundred medicines; most patients can safely stop taking medicines they no longer require. Limitations of this project include lack of overall costs of providing this service, the impact on longer term outcomes (e.g. hospitalisations) and the assumption that savings from this project will mirror pilot data; these data are being collected for future analysis.

In both systems, the bacteriocin activity decreased more quickly in the presence of wt than in the presence of LMGel, i.e. 24 h sooner in the broth culture and 1 week sooner in the meat system. In broth, strain LMG grew fastest (μmax=0.52), followed by LMGel (0.32) and finally wt (0.22). As the bacteriocin level increased initially at the same rate and reached the same peak value in the LMGel and wt cultures,

it would seem that LMGel is a somewhat less efficient bacteriocin producer than wt, possibly because of plasmid instability. As expected, bacteriocin-degrading proteolytic activity remained low (at about 7.5 U mL−1) in broth cultures seeded with LMG or Histone Methyltransferase inhibitor LMGel. In cultures seeded with wt or mt, it was about twice as high at the start of the culture and seven times as high at the end. Here, we demonstrate that sakacin P production in L. curvatus is encoded by a plasmid. This is clear from the nonbacteriocinogenic phenotype of our plasmid-cured mutants, from binding of a sakacin-specific probe to plasmid restriction fragments on Southern blots, and from binding of the same

probe to the amplicon produced by PCR from gel-purified plasmid DNA. This approximately 10-kb plasmid, furthermore, appears to exist in two different forms and additionally Seliciclib purchase to confer streptomycin resistance and the ability to ferment d-celobiose, gentiobiose, and N-acetylglucosamine. These extra features facilitate selection of plasmid-containing cells (with streptomycin) or offer a means of enhancing plasmid

stability (using carbohydrates that can be fermented only if the plasmid is present). Plasmids associated with bacteriocin production, antibiotic resistance, and/or metabolic traits are common in lactobacilli (reviewed in Wang & Lee, 1997). Sometimes, the bacteriocin-encoding plasmid Bacterial neuraminidase also carries a gene conferring immunity to the bacteriocin concerned, and plasmid loss results in sensitivity to that bacteriocin (Møretrøet al., 2005). Here, our plasmid-cured mt isolates remained resistant to sakacin P (plate assays and high-titre exposure, data not shown). This contrasts with the situation described by Møretrøet al. (2005). Naturally occurring LAB have long been used in food technology, and the use of genetically engineered LAB with improved features is envisaged for many applications. In the European Union, regulation EC1829/2003 prohibits the use of genetically modified organisms in human food unless the proposed use has been approved on the basis of evidence that it is safe for human health, animal health, and the environment and that it neither misleads the consumer nor diminishes the nutritional quality of the food. This regulation is applicable to strain LMGel. Regarding the safety of this strain, it is worth stressing that this is genetic engineering performed between two LAB strains of the same genus that are suitable for use in food.

These organisms use diverse biochemical mechanisms, such as Kodama and 4S pathways, to metabolize various polyaromatic sulfur heterocycles (PASHs). Of these, R. erythropolis IGTS8 was the first to be isolated for its ability to specifically cleave the C–S bond in PASHs without affecting the C–C bond (Kilbane & Jackowski, 1992). Since then, several Rhodococcus strains INCB024360 clinical trial have been studied (Izumi et al., 1994; Li et al., 1996; Ohshiro et al., 1996; Honda et al., 1998; Davoodi-Dehaghani et al., 2010) for specifically desulfurizing DBT and its derivatives via the 4S pathway. The desulfurization rates exhibited by

the wild-type bacteria are too low for commercialization (Kilbane, 2006). Despite numerous experimental efforts including genetic manipulations, desirable desulfurization rates are yet to be attained. From our study, it seems that this may be due to the fact that most of these studies have solely targeted the 4S pathways and desulfurizing (dsz) genes. Because the cellular phenotypes are the manifestations check details of complex interactions among various gene products and environmental factors, a systems biology

approach is critical for studying desulfurization. A comprehensive modeling approach can complement the existing and future experimental studies considerably. Such an approach would facilitate a more quantitative and insightful understanding of the interdependencies among the various pathways and associated reactions that largely determine the metabolic fluxes within

a desulfurizing strain, and hence its desulfurization activity. The resulting knowledge can then guide the design of environment and re-engineering of strains for enhancing desulfurization via the 4S pathway. This work represents the first attempt, to our knowledge, at reconstructing a stoichiometric model for the sulfur metabolism in R. erythropolis. It comprises a network of reaction Adenosine pathways involved in sulfur and central metabolism, and quantitatively describes the assimilation of sulfur from different sources into various biomass precursors. It successfully predicts two independent cell growth data and several phenotypes reported in the literature such as the effects of sulfate and various carbon sources on biodesulfurization activity. We have successfully used the model to compare the effects of eight carbon sources (citrate, ethanol, fructose, gluconate, glucose, glutamate, glycerol, and lactate) on desulfurizing activity and cell growth. The flux-based models have been widely used to study the metabolic networks of various microorganisms in a holistic manner (Burgard & Maranas, 2003; Suthers et al., 2009; Orth et al., 2010; Thiele & Palsson, 2010). Such a model for an organism is built on the known and hypothesized reactions that may take place within the organism (Gonzalez et al., 2008) based on its genomic, biochemical, and physiological information (Park et al., 2009).

These organisms use diverse biochemical mechanisms, such as Kodama and 4S pathways, to metabolize various polyaromatic sulfur heterocycles (PASHs). Of these, R. erythropolis IGTS8 was the first to be isolated for its ability to specifically cleave the C–S bond in PASHs without affecting the C–C bond (Kilbane & Jackowski, 1992). Since then, several Rhodococcus strains selleck have been studied (Izumi et al., 1994; Li et al., 1996; Ohshiro et al., 1996; Honda et al., 1998; Davoodi-Dehaghani et al., 2010) for specifically desulfurizing DBT and its derivatives via the 4S pathway. The desulfurization rates exhibited by

the wild-type bacteria are too low for commercialization (Kilbane, 2006). Despite numerous experimental efforts including genetic manipulations, desirable desulfurization rates are yet to be attained. From our study, it seems that this may be due to the fact that most of these studies have solely targeted the 4S pathways and desulfurizing (dsz) genes. Because the cellular phenotypes are the manifestations MAPK inhibitor of complex interactions among various gene products and environmental factors, a systems biology

approach is critical for studying desulfurization. A comprehensive modeling approach can complement the existing and future experimental studies considerably. Such an approach would facilitate a more quantitative and insightful understanding of the interdependencies among the various pathways and associated reactions that largely determine the metabolic fluxes within

a desulfurizing strain, and hence its desulfurization activity. The resulting knowledge can then guide the design of environment and re-engineering of strains for enhancing desulfurization via the 4S pathway. This work represents the first attempt, to our knowledge, at reconstructing a stoichiometric model for the sulfur metabolism in R. erythropolis. It comprises a network of reaction selleck kinase inhibitor pathways involved in sulfur and central metabolism, and quantitatively describes the assimilation of sulfur from different sources into various biomass precursors. It successfully predicts two independent cell growth data and several phenotypes reported in the literature such as the effects of sulfate and various carbon sources on biodesulfurization activity. We have successfully used the model to compare the effects of eight carbon sources (citrate, ethanol, fructose, gluconate, glucose, glutamate, glycerol, and lactate) on desulfurizing activity and cell growth. The flux-based models have been widely used to study the metabolic networks of various microorganisms in a holistic manner (Burgard & Maranas, 2003; Suthers et al., 2009; Orth et al., 2010; Thiele & Palsson, 2010). Such a model for an organism is built on the known and hypothesized reactions that may take place within the organism (Gonzalez et al., 2008) based on its genomic, biochemical, and physiological information (Park et al., 2009).

3a). Because gingipain activity can be regulated at the transcriptional and post-transcriptional levels (Tokuda et al., 1998), oligonucleotide primers, as described previously

(Vanterpool et al., 2005a), were used in RT-PCR analysis to determine whether these two sigma factors were involved in the transcriptional regulation of gingipain-encoding Selleckchem Neratinib genes. As shown in Fig. 3b, the inactivation of PG0162 and PG1660 had no effect on the expression of rgpA, rgpB, or kgp at the transcriptional level. In FLL355 (PG1827∷ermF), the Kgp activity showed a 25% increase over the wild type. No change was observed in the transcription of the kgp gene in FLL355 (data not shown). Taken together, these results suggest that ECF sigma factors may be involved in the post-transcriptional regulation of gingipains. Post-transcriptional regulation of the gingipains in P. gingivalis is associated with its maturation pathway, which is linked to the biosynthesis ATR inhibitor of surface carbohydrates (Shoji et al., 2002; Paramonov et al., 2005) and several other proteins including the PorR (Shoji et al., 2002), PorT (Sato et al., 2005; Nguyen et al., 2009),

Sov (Saiki & Konishi, 2010), and VimA (Vanterpool et al., 2006). It is unclear how these factors are modulated by the ECF sigma factors and is an active area of further exploration in the laboratory. The correlation between gingipain activity and hemagglutination in P. gingivalis (Lewis et al., 1999; Shi et al., 1999; Vanterpool et al., 2005a) is related to the similar adhesion domains encoded by the hagA, rgpA, and kgp genes (Chen & Duncan, 2004). The hemagglutination potential of ECF sigma factor-defective mutants was assessed. In comparison with the wild-type strain, there was a decrease Exoribonuclease in the hemagglutination activity in all the mutants. In FLL350, the level of hemagglutination activity was comparable

to the negative control. This is in contrast to FLL354, which showed the greatest reduction in gingipain activity, but a higher hemagglutination activity. RT-PCR using hagA-specific primers indicated no change in the expression of that gene in FLL350 (Fig. 4c). While gingipains have been observed to have hemolytic activity (Shah & Gharbia, 1989; Lewis et al., 1999), hemolysin can be independent of their catalytic association (Deshpande & Khan, 1999). Several putative hemolysin genes have been identified in the P. gingivalis genome (Nelson et al., 2003) and cloned in E. coli (Karunakaran & Holt, 1993). The hemolysins produced by P. gingivalis provide the bacterium with heme-containing molecules that are required for their in vivo survival. Hemolytic activities of all the ECF-defective mutants in this study were similar to those of the wild type, except for FLL350 (Fig. 4d). The FLL350 mutant showed a 50% reduction in those activities compared with the parent strain.

tuberculosis have been synthesized (Lilienkampf

et al., 2009; Upadhayaya et al., 2009). Diarylquinolines were also shown to kill dormant M. tuberculosis as effectively as replicating bacilli and to inhibit ATP synthesis in dormant M. smegmatis (Koul et al., 2008). This unique dual bactericidal activity, with equal potency on replicating and dormant bacilli, distinguishes diarylquinolines from all the currently used antituberculosis drugs, such as isoniazid and rifampicin. Selleck INCB024360 These front-line drugs show significantly less activity on dormant mycobacteria as compared with replicating bacilli (Koul et al., 2008; Rao et al., 2008). Thus, although ATP synthase is significantly downregulated during dormancy, its residual activity Osimertinib price appears to be essential for mycobacteria irrespective of their

physiological state. This makes ATP synthase an efficient drug target to tackle both replicating as well as dormant bacilli. In vivo experiments using mouse models indicated that diarylquinolines have bactericidal activity exceeding the effect of current first-line antibiotics (Andries et al., 2005; Lounis et al., 2006). Diarylquinolines, in particular when applied in combination therapy with the first-line antibiotic pyrazinamide, have a strong potential for shortening the duration of tuberculosis treatment (Lounis et al., 2006; Ibrahim et al., 2007). The physiological basis for this observed synergy remains obscure. In phase IIb clinical tests, the addition of TMC207 to standard therapy strongly decreased the count of CFU in the sputum of patients with multi-drug-resistant tuberculosis as compared find more with an active-placebo group (Diacon et al., 2009). TMC207 also accelerated conversion to a negative sputum culture, as compared with a placebo. These findings validate ATP synthase as a target for the treatment of tuberculosis. Respiratory ATP production is not only essential for growth, but also represents a critical weak point in dormant mycobacteria. Although most enzymes involved in respiratory

ATP synthesis are conserved between prokaryotes and eukaryotes, targeting ATP production may be a highly efficient approach for the development of antibacterial drugs. The strategy may be to target enzymes, which do not have homologs in human metabolism, as in the case of NDH-2. Alternatively, as applied for ATP synthase, small differences in the structure between a bacterial enzyme and a human homologue may be utilized for selective inhibition. Understanding respiratory ATP production in replicating and dormant mycobacteria will not only fuel development of novel drugs but also shed light on how these bacteria perform their intriguing task of extreme persistence without significant growth. The authors wish to thank Prof. Dr H. Lill (VU Amsterdam) and Prof. Dr K. Andries (Johnson and Johnson) for critically reading the manuscript, and Dr J. Guillemont, Dr E.

, 2009a). The apical endocytic recycling selleck model in filamentous fungi has been widely accepted (Steinberg, 2007; Taheri-Talesh et al., 2008; Upadhyay & Shaw, 2008; Abenza et al., 2009; Peñalva, 2010). Notably, in

A. oryzae, endocytosis-deficient hyphae display severe defective growth, suggesting that endocytosis and apical growth are highly linked (Higuchi et al., 2009b). In Aspergillus nidulans, similar localization and functional analyses of endocytic proteins, such as AbpA, AmpA, FimA, and SlaB, the orthologs of S. cerevisiae Abp1p, Rvs167p, Sac6p, and End4p/Sla2p, respectively, have been reported (Araujo-Bazán et al., 2008; Taheri-Talesh et al., 2008; Upadhyay & Shaw, 2008; Hervas-Aguilar & Penalva, 2010). Although novel insights,

Trametinib solubility dmso such as apical endocytic recycling, have been elucidated based on the analyses of ortholog proteins of S. cerevisiae, a more detailed mechanism related to endocytosis in the hyphal tip region has not yet been clarified (Peñalva, 2010). AAA ATPases are conserved from prokaryotes to humans and play roles in various processes such as membrane fusion and protein degradation (White & Lauring, 2007). AAA ATPases contain the ATPase domain at the C-terminus with high homology, but the rest of the regions are not well conserved. Moreover, AAA ATPases form a ring-shaped hexamer with a central pore and the ATPase domain facing inside. In the endocytic pathway, an AAA ATPase Vps4p in S. cerevisiae functions in the disassembly of the ESCRT (endosomal sorting complexes required for transport)-III for complex from the membrane of multivesicular bodies (MVBs) to the cytoplasm (Babst et al., 1997, 1998; Saksena et al., 2009). Although there are several reports on AAA ATPases in many organisms, no protein that functions in endocytosis has been reported so far (White & Lauring, 2007). According to the analyses of endocytic proteins in A. oryzae, the mechanism of endocytosis, which

is characteristic of the organism, seems to exist at the apical region. We, therefore, explored novel components associated with endocytosis by yeast two-hybrid (YTH) screening using an endocytic marker protein AoAbp1, having two SH3 domains at the C-terminal region, which are related to endocytic protein–protein interaction, as bait. Of the candidates obtained by YTH screening, the gene aipA was found, which likely encodes AAA ATPase. The interaction between AipA and AoAbp1 by YTH and in vitro, in vivo localization, and functional analyses using the aipA-overexpressing strain suggested that AipA is a putative AAA ATPase negatively functioning in apical endocytic recycling. The A. oryzae strains used in this study are listed in Table 1. The DNA cloning methods used in this study were described previously (Higuchi et al., 2009b). All plasmids used for A. oryzae transformation in this study were constructed by the MultiSite Gateway System (Invitrogen).