Genetically altering the disease vector status of insects using recombinant DNA technologies is being considered as an alternative to eradication efforts. Manipulating endogenous immune response mosquitoes such temporal and special expression antimicrobial peptides like cecropin may result in a refractory phenotype. Using transgenic technology unique pattern A (cecA) Anopheles gambiae was created that cecA expressed beginning 24 h after blood meal posterior midgut. Two independent lines An....

Eukaryotic genomes are organized into chromatin domains with three-dimensional arrangements that presumably result from interactions between the constituents—proteins, DNA, and RNA—within physical constraints of nucleus. We used chromosome conformation capture (3C) followed by high-throughput sequencing (Hi-C) wild-type mutant strains Neurospora crassa to gain insight role heterochromatin in organization function genome. tested three proteins thought be important for establishment...

Significance Two forms of heterochromatin, constitutive and facultative, cause gene silencing in eukaryotes. In Neurospora crassa , H3K27me2/3-marked facultative heterochromatin reversibly represses scores specialized genes, whereas H3K9me3-marked permanently silences repetitive DNA. Interactions between provide a structural framework for the genome, this is thought to be functionally important. Histone marks underlying are nonessential N. permitting tests their roles genome organization...

Summary 6S RNA binds σ 70 ‐RNA polymerase and downregulates transcription at many ‐dependent promoters, but others escape regulation even during stationary phase when the majority of machinery is bound by RNA. We report that core promoter elements determine this specificity; a weak −35 element allows to be sensitive, an extended −10 similarly determines inhibition except consensus present. These two features together predicted hundreds mapped Escherichia coli promoters might subject...

Comparative genomics has recently provided unprecedented insights into the biology and evolution of fungal lineage. In postgenomics era, a major research interest focuses now on detailing functions genomes, i.e. how genomic information manifests complex phenotypes. Emerging evidence across diverse eukaryotes revealed that organization DNA within nucleus is critically important. Here, we discuss current knowledge genome organization, from association chromosomes to topological structures at...

Abstract Polycomb group (PcG) proteins form chromatin modifying complexes that stably repress lineage- or context-specific genes in animals, plants, and some fungi. Repressive Complex 2 (PRC2) catalyzes trimethylation of lysine 27 on histone H3 (H3K27me3) to assemble repressive chromatin. In the model fungus Neurospora crassa, H3K27me3 deposition is controlled by H3K36 methyltransferase ASH1 components constitutive heterochromatin including H3K9me3-binding protein HETEROCHROMATIN PROTEIN 1...

Significance Regulatory networks change during evolution. A protein that controls many genes in one species may control a different set of another. We examined how mRNA evolve, focusing on the PUF (Pumilio and FBF) family RNA-binding proteins. These govern stability translation hundreds mRNAs enable coordinate regulation discrete biological outcomes. To understand RNA we used knowledge specificity each to predict its targets directly identified bound divergent fungi via biochemical methods....

Abstract The eukaryotic genome must be precisely organized for its proper function, as topology impacts transcriptional regulation, cell division, replication, and repair, among other essential processes. Disruptions to human can lead diseases, including cancer. advent of chromosome conformation capture with high-throughput sequencing (Hi-C) assess organization has revolutionized the study nuclear topology; Hi-C elucidated numerous genomic structures, chromosomal territories, active/silent...

Summary 6S RNA is a small, non‐coding that interacts with σ 70 ‐RNA polymerase and downregulates transcription at many promoters during stationary phase. When bound to polymerase, engaged in the active site of manner similar enough promoter DNA can serve as template for synthesis. It has been proposed mimics conformation initiation, suggesting contacts between or may be similar. Here we demonstrate region 4.2 critical interaction polymerase. We define an expanded binding surface encompasses...

Heterochromatin and associated gene silencing processes play roles in development, genome defense, chromosome function. In many species, constitutive heterochromatin is decorated with histone H3 tri-methylated at lysine 9 (H3K9me3) cytosine methylation. Neurospora crassa, a five-protein complex, DCDC, catalyzes H3K9 methylation, which then directs DNA Here, we identify characterize important for DCDC function, dim-3 (defective methylation-3), encodes the nuclear import chaperone NUP-6...

Summary Mismatch repair (MMR) corrects DNA polymerase errors occurring during genome replication. MMR is critical for maintenance, and its loss increases mutation rates several hundred fold. Recent work has shown that the interaction between mismatch recognition protein MutS replication processivity clamp important in Bacillus subtilis . To further understand how coupled to replication, we examined subcellular localization of proteins fused green fluorescent (GFP) live cells, following an...

ABSTRACT The β clamp is an essential replication sliding required for processive DNA synthesis. also critical several additional aspects of metabolism, including mismatch repair (MMR). dnaN5 allele Bacillus subtilis encodes a mutant form containing the G73R substitution. Cells with are temperature sensitive growth due to defect in at 49°C, and they show increase mutation frequency caused by partial MMR permissive temperatures. We selected intragenic suppressors that rescued viability 49°C...

In the filamentous fungus Neurospora crassa, constitutive heterochromatin is marked by tri-methylation of histone H3 lysine 9 (H3K9me3) and DNA methylation. We identified mutations in defective methylation-1 (dim-1) gene that cause defects cytosine methylation implicate a putative AAA-ATPase chromatin remodeler. Although it was well-established remodelers can affect transcription influencing accessibility with nucleosomes, little known about role on normally not transcribed, including...

DNA methylation, a prototypical epigenetic modification implicated in gene silencing, occurs many eukaryotes and plays significant role the etiology of diseases such as cancer. The filamentous fungus

Chromosomes must correctly fold in eukaryotic nuclei for proper genome function. Eukaryotic organisms hierarchically organize their genomes, including the fungus Neurospora crassa , where chromatin fiber loops compact into Topologically Associated Domain-like structures formed by heterochromatic region aggregation. However, insufficient data exist on how histone posttranslational modifications (PTMs), acetylation, affect organization. In Neurospora, HCHC complex [composed of proteins HDA-1,...

Both prokaryotes and eukaryotes respond to DNA damage through a complex set of physiological changes. Alterations in gene expression, the redistribution existing proteins, assembly new protein complexes can be stimulated by variety lesions mismatched base pairs. Fluorescence microscopy has been used as powerful experimental tool for visualizing quantifying these other responses monitor replication status within subcellular architecture living cell. Translational fusions between fluorescent...

Genome organization is essential for proper function, including gene expression. In metazoan genome organization, chromatin loops and Topologically Associated Domains (TADs) facilitate local clustering, while chromosomes form distinct nuclear territories characterized by compartmentalization of silent heterochromatin at the periphery active euchromatin in nucleus center. A similar hierarchical occurs fungus

Organization of the eukaryotic genome is essential for proper function, including gene expression. In metazoans, chromatin loops and Topologically Associated Domains (TADs) organize genes into transcription factories, while chromosomes occupy nuclear territories in which silent heterochromatin compartmentalized at periphery active euchromatin localizes to nucleus center. A similar hierarchical organization occurs fungus Neurospora crassa where its seven form a Rabl conformation typified by...

Chromosomes must correctly fold in eukaryotic nuclei for proper genome function. Eukaryotic organisms hierarchically organize their genomes, including the fungus Neurospora crassa, where chromatin fiber loops compact into Topologically Associated Domain (TAD)-like structures formed by heterochromatic region aggregation. However, insufficient data exists on how histone post-translational modifications, acetylation, affect organization. In Neurospora, HCHC complex (comprised of proteins HDA-1,...