Cellular remodeling and JAK inhibition promote zygotic gene expression in the Ciona germline

During development, remodeling of the cellular transcriptome and proteome underlies cell fate decisions and, in somatic lineages, transcription control is a major determinant of fateful biomolecular transitions. By contrast, early germline fate specification in numerous vertebrate and invertebrate species relies extensively on RNA-level regulation, exerted on asymmetrically inherited maternal supplies, with little-to-no zygotic transcription. However delayed, a maternal-to-zygotic transition is nevertheless poised to complete the deployment of pre-gametic programs in the germline. Here, we focused on early germline specification in the tunicate Ciona to study zygotic genome activation. We first demonstrate that a peculiar cellular remodeling event excludes localized postplasmic mRNAs, including Pem-1, which encodes the general inhibitor of transcription. Subsequently, zygotic transcription begins in Pem-1-negative primordial germ cells (PGCs), as revealed by histochemical detection of elongating RNA Polymerase II (RNAPII), and nascent transcripts from the Mef2 locus. Using PGC-specific Mef2 transcription as a read-out, we uncovered a provisional antagonism between JAK and MEK/BMPRI/GSK3 signaling, which controls the onset of zygotic gene expression, following cellular remodeling of PGC progenitor cells. We propose a 2-step model for the onset of zygotic transcription in the Ciona germline, which relies on successive cellular remodeling and JAK inhibition, and discuss the significance of germ plasm dislocation and remodeling in the context of developmental fate specification.

Chemical Signals are Involved in the Detection and Preference of Food Sources in Blattella Germanica

The German cockroach (Blattella germanica) is a cosmopolitan omnivorous species which can take nutrients from a wide variety of sources. Its condition of pest generated interest in the study of its biology, and while there are many works regarding its feeding behavior, few approaches have been made to the relevance of the chemosensory system during foraging. Objective: In this work we studied the role of chemical stimuli in the detection and preference of food sources in B. germanica using behavioral observations. Methods: Adult males were placed in a circular experimental arena where different diet types were presented individually. Parameters associated to the detection of odors (latency to the food source, number of visits to the source) were measured in an experimental time of 15 minutes. We also measured the total amount ingested determining the weight gain of each individual, as an indicator of taste evaluation of food. Findings: Insects showed a lower latency when the source emitted a larger amount of volatile compounds, so peanut butter was detected faster compared with the other diets studied. Regarding ingestion, insects spent significantly more time on the peanut butter, indicating the presence of phagostimulants such as sugars or lipids. When sugar solutions were used as food sources, the number of visits, the time spent on the source and the amount consumed was higher when increasing sugar concentration. Insects also showed preference for the solution of higher concentration, when two sugar solutions were presented simultaneously. Latency increased and the amount of sugar ingested decreased when insects´ mouthparts were chemically blocked with N-Ethylmaleimide, a general inhibitor of chemoreception. Novelty/Improvement: These results describe how cockroaches use odors to locate food sources and chemoreceptors in their mouthparts to identify diets of higher energetic value, from information regarding the concentration and quality of the food source. Understanding the feeding behavior of cockroaches allows the design of more attractive and palatable toxic gel baits enhancing their efficacy for cockroach control.

SLFL Genes Participate in the Ubiquitination and Degradation ofS-RNase in Self-Compatible Chinese Peach

The gametophytic self-incompatibility (SI) mediated by S-RNase of Rosaceae, Solanaceae and Plantaginaceae, is controlled by two tightly linked genes located at highly polymorphic S-locus: the S-RNase for pistil specificity and the F-box gene (SFB/SLF) for pollen specificity, respectively. The F-box gene of peach (Prunus persica) isShaplotype-specific F-box (SFB). In this study, we selected 37 representative varieties according to the evolution route of peach and identified their S genotypes. We cloned pollen determinant genes mutantPperSFB1m, PperSFB2m, PperSFB4mand normalPperSFB2, and style determinant genesS1-RNase, S2-RNase, S2m-RNaseandS4-RNase.MutantPperSFBswere translated terminated prematurely because of fragment insertion. Yeast two-hybrid showed that mutant PperSFBs and normal PperSFB2 interacted with all S-RNases. NormalPperSFB2was divided into four parts: box, box-V1, V1-V2 and HVa-HVb. Protein interaction analyses showed that the box portion did not interact with S-RNases, both of the box-V1 and V1-V2 had interactions with S-RNases, while the hypervariable region ofPperSFB2HVa-HVb only interacted with S2-RNase. Bioinformatics analysis of peach genome revealed that there were other F-box genes located at S-locus, and of which three F-box genes were specifically expressed in pollen, namelyPperSLFL1, PperSLFL2andPperSLFL3, respectively. Phylogenetic analysis showed that PperSFBs and PperSLFLs were classified into two different clades. Yeast two-hybrid analysis revealed that as with PperSFBs, the three F-box proteins interacted with PperSSK1. Yeast two-hybrid and BiFC showed that PperSLFLs interacted with S-RNases with no allelic specificity. In vitro ubiquitination assay showed that PperSLFLs could tag ubiquitin molecules to PperS-RNases. In all, the above results suggest that threePperSLFLsare the appropriate candidates for the ‘general inhibitor’, which would inactivate the S-RNases in pollen tubes, and the role of three PperSLFL proteins is redundant, as S-RNase repressors involved in the self-incompatibility of peach.

p58IPK is an inhibitor of the eIF2α kinase GCN2 and its localization and expression underpin protein synthesis and ER processing capacity

We show that p58IPK is a general inhibitor of the eIF2α kinases and its expression and localization are important in the capacity of the cells to respond to cellular stress by controlling protein synthesis rates and subsequent folding in the endoplasmic reticulum.

Indole inhibition of N-acylated homoserine lactone-mediated quorum signalling is widespread in Gram-negative bacteria

The LuxI/R quorum-sensing system and its associated N-acylated homoserine lactone (AHL) signal is widespread among Gram-negative bacteria. Although inhibition by indole of AHL quorum signalling in Pseudomonas aeruginosa and Acinetobacter oleivorans has been reported previously, it has not been documented among other species. Here, we show that co-culture with wild-type Escherichia coli, but not with E. coli tnaA mutants that lack tryptophanase and as a result do not produce indole, inhibits AHL-regulated pigmentation in Chromobacterium violaceum (violacein), Pseudomonas chlororaphis (phenazine) and Serratia marcescens (prodigiosin). Loss of pigmentation also occurred during pure culture growth of Chro. violaceum, P. chlororaphis and S. marcescens in the presence of physiologically relevant indole concentrations (0.5–1.0 mM). Inhibition of violacein production by indole was counteracted by the addition of the Chro. violaceum cognate autoinducer, N-decanoyl homoserine lactone (C10-HSL), in a dose-dependent manner. The addition of exogenous indole or co-culture with E. coli also affected Chro. violaceum transcription of vioA (violacein pigment production) and chiA (chitinase production), but had no effect on pykF (pyruvate kinase), which is not quorum regulated. Chro. violaceum AHL-regulated elastase and chitinase activity were inhibited by indole, as was motility. Growth of Chro. violaceum was not affected by indole or C10-HSL supplementation. Using a nematode-feeding virulence assay, we observed that survival of Caenorhabditis elegans exposed to Chro. violaceum, P. chlororaphis and S. marcescens was enhanced during indole supplementation. Overall, these studies suggest that indole represents a general inhibitor of AHL-based quorum signalling in Gram-negative bacteria.