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https://www.readbyqxmd.com/read/27922451/direct-screening-for-chromatin-status-on-dna-barcodes-in-yeast-delineates-the-regulome-of-h3k79-methylation-by-dot1
#1
Hanneke Vlaming, Thom M Molenaar, Tibor van Welsem, Deepani W Poramba-Liyanage, Desiree E Smith, Arno Velds, Liesbeth Hoekman, Tessy Korthout, Sjoerd Hendriks, Af Maarten Altelaar, Fred van Leeuwen
Given the frequent misregulation of chromatin in cancer, it is important to understand the cellular mechanisms that regulate chromatin structure. However, systematic screening for epigenetic regulators is challenging and often relies on laborious assays or indirect reporter read-outs. Here we describe a strategy, Epi-ID, to directly assess chromatin status in thousands of mutants. In Epi-ID, chromatin status on DNA barcodes is interrogated by chromatin immunoprecipitation followed by deep sequencing, allowing for quantitative comparison of many mutants in parallel...
December 6, 2016: ELife
https://www.readbyqxmd.com/read/27903907/chromatin-binding-of-gcn5-in-drosophila-is-largely-mediated-by-cp190
#2
Tamer Ali, Marcus Krüger, Sabin Bhuju, Michael Jarek, Marek Bartkuhn, Rainer Renkawitz
Centrosomal 190 kDa protein (CP190) is a promoter binding factor, mediates long-range interactions in the context of enhancer-promoter contacts and in chromosomal domain formation. All Drosophila insulator proteins bind CP190 suggesting a crucial role in insulator function. CP190 has major effects on chromatin, such as depletion of nucleosomes, high nucleosomal turnover and prevention of heterochromatin expansion. Here, we searched for enzymes, which might be involved in CP190 mediated chromatin changes. Eighty percent of the genomic binding sites of the histone acetyltransferase Gcn5 are colocalizing with CP190 binding...
November 29, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27899560/not5-dependent-co-translational-assembly-of-ada2-and-spt20-is-essential-for-functional-integrity-of-saga
#3
Sari Kassem, Zoltan Villanyi, Martine A Collart
Acetylation of histones regulates gene expression in eukaryotes. In the yeast Saccharomyces cerevisiae it depends mainly upon the ADA and SAGA histone acetyltransferase complexes for which Gcn5 is the catalytic subunit. Previous screens have determined that global acetylation is reduced in cells lacking subunits of the Ccr4-Not complex, a global regulator of eukaryotic gene expression. In this study we have characterized the functional connection between the Ccr4-Not complex and SAGA. We show that SAGA mRNAs encoding a core set of SAGA subunits are tethered together for co-translational assembly of the encoded proteins...
November 28, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27894818/supt20-is-required-for-development-of-the-axial-skeleton
#4
Sunita Warrier, Samer Nuwayhid, Julia A Sabatino, Kelsey F Sugrue, Irene E Zohn
Somitogenesis and subsequent axial skeletal development is regulated by the interaction of pathways that determine the periodicity of somite formation, rostrocaudal somite polarity and segment identity. Here we use a hypomorphic mutant mouse line to demonstrate that Supt20 (Suppressor of Ty20) is required for development of the axial skeleton. Supt20 hypomorphs display fusions of the ribs and vertebrae at lower thoracic levels along with anterior homeotic transformation of L1 to T14. These defects are preceded by reduction of the rostral somite and posterior shifts in Hox gene expression...
November 25, 2016: Developmental Biology
https://www.readbyqxmd.com/read/27876471/cobalamin-and-folate-protect-mitochondrial-and-contractile-functions-in-a-murine-model-of-cardiac-pressure-overload
#5
Jérôme Piquereau, Maryline Moulin, Giada Zurlo, Philippe Mateo, Mélanie Gressette, Jean-Louis Paul, Christophe Lemaire, Renée Ventura-Clapier, Vladimir Veksler, Anne Garnier
PGC-1α, a key regulator of energy metabolism, seems to be a relevant therapeutic target to rectify the energy deficit observed in heart failure (HF). Since our previous work has shown positive effects of cobalamin (Cb) on PGC-1α cascade, we investigate the protective role of Cb in pressure overload-induced myocardial dysfunction. Mice were fed with normal diet (ND) or with Cb and folate supplemented diet (SD) 3weeks before and 4weeks after transverse aortic constriction (TAC). At the end, left ventricle hypertrophy and drop of ejection fraction were significantly lower in SD mice than in ND mice...
November 19, 2016: Journal of Molecular and Cellular Cardiology
https://www.readbyqxmd.com/read/27874008/the-gcn5-cited2-pka-signalling-module-controls-hepatic-glucose-metabolism-through-a-camp-induced-substrate-switch
#6
Mashito Sakai, Tomoko Tujimura-Hayakawa, Takashi Yagi, Hiroyuki Yano, Masaru Mitsushima, Hiroyuki Unoki-Kubota, Yasushi Kaburagi, Hiroshi Inoue, Yoshiaki Kido, Masato Kasuga, Michihiro Matsumoto
Hepatic gluconeogenesis during fasting results from gluconeogenic gene activation via the glucagon-cAMP-protein kinase A (PKA) pathway, a process whose dysregulation underlies fasting hyperglycemia in diabetes. Such transcriptional activation requires epigenetic changes at promoters by mechanisms that have remained unclear. Here we show that GCN5 functions both as a histone acetyltransferase (HAT) to activate fasting gluconeogenesis and as an acetyltransferase for the transcriptional co-activator PGC-1α to inhibit gluconeogenesis in the fed state...
November 22, 2016: Nature Communications
https://www.readbyqxmd.com/read/27816487/arylalkylamine-n-acetyltransferase-1-gene-tcaanat1-is-required-for-cuticle-morphology-and-pigmentation-of-the-adult-red-flour-beetle-tribolium-castaneum
#7
Mi Young Noh, Bonwoo Koo, Karl J Kramer, Subbaratnam Muthukrishnan, Yasuyuki Arakane
In the insect cuticle tanning pathway (sclerotization and pigmentation), the enzyme arylalkylamine N-acetyltransferase (AANAT) catalyzes the acetylation of dopamine to form N-acetyldopamine (NADA), which is one of the major precursors for quinone-mediated tanning. In this study we characterized and investigated the function of TcAANAT1 in cuticle pigmentation of the red flour beetle, Tribolium castaneum. We isolated a full length TcAANAT1 cDNA that encodes a protein of 256 amino acid residues with a predicted GCN5-related acetyltransferase domain containing an acetyl-CoA binding motif...
November 2, 2016: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/27803161/molecular-basis-for-cohesin-acetylation-by-establishment-of-sister-chromatid-cohesion-n-acetyltransferase-esco1
#8
Yadilette Rivera-Colón, Andrew Maguire, Glen P Liszczak, Adam S Olia, Ronen Marmorstein
Protein acetylation is a prevalent posttranslational modification that is regulated by diverse acetyltransferase enzymes. While histone acetyltransferases (HATs) have been well characterized both structurally and mechanistically, far less is known about non-histone acetyltransferase enzymes. The human ESCO1 and ESCO2 paralogs acetylate the cohesin complex subunit SMC3 to regulate the separation of sister chromatids during mitosis and meiosis. Missense mutations within the acetyltransferase domain of these proteins correlate with diseases, including endometrial cancers and Roberts Syndrome...
November 1, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27796307/kat2a-kat2b-targeted-acetylome-reveals-a-role-for-plk4-acetylation-in-preventing-centrosome-amplification
#9
Marjorie Fournier, Meritxell Orpinell, Cédric Grauffel, Elisabeth Scheer, Jean-Marie Garnier, Tao Ye, Virginie Chavant, Mathilde Joint, Fumiko Esashi, Annick Dejaegere, Pierre Gönczy, László Tora
Lysine acetylation is a widespread post-translational modification regulating various biological processes. To characterize cellular functions of the human lysine acetyltransferases KAT2A (GCN5) and KAT2B (PCAF), we determined their acetylome by shotgun proteomics. One of the newly identified KAT2A/2B substrate is polo-like kinase 4 (PLK4), a key regulator of centrosome duplication. We demonstrate that KAT2A/2B acetylate the PLK4 kinase domain on residues K45 and K46. Molecular dynamics modelling suggests that K45/K46 acetylation impairs kinase activity by shifting the kinase to an inactive conformation...
October 31, 2016: Nature Communications
https://www.readbyqxmd.com/read/27785912/structural-and-biochemical-characterization-of-acinetobacter-spp-aminoglycoside-acetyltransferases-highlights-functional-and-evolutionary-variation-among-antibiotic-resistance-enzymes
#10
Peter J Stogios, Misty Lee Kuhn, Elena Evdokimova, Melissa Law, Patrice Courvalin, Alexei Savchenko
Modification of aminoglycosides by N-acetyltransferases (AACs) is one of the major mechanisms of resistance to these antibiotics in human bacterial pathogens. More than fifty enzymes belonging to the AAC(6') subfamily have been identified in Gram-negative and Gram-positive clinical isolates. Our understanding of the molecular function and evolutionary origin of these resistance enzymes remains incomplete. Here we report the structural and enzymatic characterization of AAC(6')-Ig and AAC(6')-Ih from Acinetobacter spp...
October 27, 2016: ACS Infectious Diseases
https://www.readbyqxmd.com/read/27783928/insight-into-the-3d-structure-and-substrate-specificity-of-previously-uncharacterized-gnat-superfamily-acetyltransferases-from-pathogenic-bacteria
#11
Karolina A Majorek, Tomasz Osinski, David T Tran, Alina Revilla, Wayne F Anderson, Wladek Minor, Misty L Kuhn
Members of the Gcn5-related N-acetyltransferase (GNAT) superfamily catalyze the acetylation of a wide range of small molecule and protein substrates. Due to their abundance in all kingdoms of life and diversity of their functions, they are implicated in many aspects of eukaryotic and prokaryotic physiology. Although numerous GNATs have been identified thus far, many remain structurally and functionally uncharacterized. The elucidation of their structures and functions is critical for broadening our knowledge of this diverse and important superfamily...
January 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27741413/saga-complex-and-gcn5-are-necessary-for-respiration-in-budding-yeast
#12
Claudia Canzonetta, Manuela Leo, Salvatore Rocco Guarino, Arianna Montanari, Silvia Francisci, Patrizia Filetici
In budding yeast, growth through fermentation and/or respiration is dependent on the type of carbon source present in the medium. SAGA complex is the main acetylation complex and is required, together with Rtg factors, for nucleus-mitochondria communication and transcriptional activation of specific nuclear genes. Even though acetylation is necessary for mitochondria activity and respiratory pathways the direct role of histone acetyltransferases and SAGA complex has never been investigated directly. In this study we demonstrate, for the first time, that Gcn5 and SAGA are needed for respiratory metabolism and oxygen consumption...
December 2016: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/27694804/n-acetylglucosamine-sensing-by-a-gcn5-related-n-acetyltransferase-induces-transcription-via-chromatin-histone-acetylation-in-fungi
#13
Chang Su, Yang Lu, Haoping Liu
N-acetylglucosamine (GlcNAc) exists ubiquitously as a component of the surface on a wide range of cells, from bacteria to humans. Many fungi are able to utilize environmental GlcNAc to support growth and induce cellular development, a property important for their survival in various host niches. However, how the GlcNAc signal is sensed and subsequently transduced is largely unknown. Here, we identify a gene that is essential for GlcNAc signalling (NGS1) in Candida albicans, a commensal and pathogenic yeast of humans...
October 3, 2016: Nature Communications
https://www.readbyqxmd.com/read/27694229/phosphinothricin-acetyltransferases-identified-using-in-vivo-in-vitro-and-bioinformatic-analyses
#14
Chelsey M VanDrisse, Kristy L Hentchel, Jorge C Escalante-Semerena
Acetylation of small molecules is widespread in nature, and in some cases, cells use this process to detoxify harmful chemicals. Streptomyces species utilize a Gcn5 N-acetyltransferase (GNAT), known as Bar, to acetylate and detoxify a self-produced toxin, phosphinothricin (PPT), a glutamate analogue. Bar homologues, such as MddA from Salmonella enterica, acetylate methionine analogues such as methionine sulfoximine (MSX) and methionine sulfone (MSO), but not PPT, even though Bar homologues are annotated as PPT acetyltransferases...
December 15, 2016: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/27690055/arbitrarily-accessible-3d-microfluidic-device-for-combinatorial-high-throughput-drug-screening
#15
Zhuofa Chen, Weizhi Li, Gihoon Choi, Xiaonan Yang, Jun Miao, Liwang Cui, Weihua Guan
Microfluidics-based drug-screening systems have enabled efficient and high-throughput drug screening, but their routine uses in ordinary labs are limited due to the complexity involved in device fabrication and system setup. In this work, we report an easy-to-use and low-cost arbitrarily accessible 3D microfluidic device that can be easily adopted by various labs to perform combinatorial assays for high-throughput drug screening. The device is capable of precisely performing automatic and simultaneous reagent loading and aliquoting tasks and performing multistep assays with arbitrary sequences...
2016: Sensors
https://www.readbyqxmd.com/read/27672091/identification-of-tension-sensing-motif-of-histone-h3-in-saccharomyces-cerevisiae-and-its-regulation-by-histone-modifying-enzymes
#16
Jianjun Luo, Xiexiong Deng, Christopher Buehl, Xinjing Xu, Min-Hao Kuo
To ensure genome stability during cell division, all chromosomes must attach to spindles emanating from the opposite spindle pole bodies before segregation. The tension between sister chromatids generated by the poleward pulling force is an integral part of chromosome bi-orientation. In budding yeast, the residue Gly44 of histone H3 is critical for retaining the conserved Shugoshin protein Sgo1p at the pericentromeres for monitoring the tension status during mitosis. Studies carried out in this work showed that Lys42, Gly44, and Thr45 of H3 form the core of a tension sensing motif, TSM...
September 26, 2016: Genetics
https://www.readbyqxmd.com/read/27647933/wnt-%C3%AE-catenin-dependent-acetylation-of-pygo2-by-cbp-p300-histone-acetyltransferase-family-members
#17
Kenneth Kao, Phillip Andrews
Pygopus 2 (Pygo2) is a chromatin effector that plays an essential role in canonical Wnt signaling associated with development and stem cell growth. Its function is to facilitate histone acetylation by recruitment of histone acetyltransferases (HATs) at active sites of β-catenin-mediated transcription. In this study, we report that Pygo2 itself, is transiently acetylated when bound to the activated TCF/β-catenin transcription complex, which correlated with β-catenin binding and Axin2 gene activation. The HATs CBP/p300, but not GCN5/PCAF targeted specific Lysine residues of the N-terminal homology domain of Pygo2 for acetylation...
September 19, 2016: Biochemical Journal
https://www.readbyqxmd.com/read/27647735/four-domains-of-ada1-form-a-heterochromatin-boundary-through-different-mechanisms
#18
Kazuma Kamata, Kaori Shinmyozu, Jun-Ichi Nakayama, Masanori Hatashita, Hiroyuki Uchida, Masaya Oki
In eukaryotic cells, there are two chromatin states, silenced and active, and the formation of a so-called boundary plays a critical role in demarcating these regions; however, the mechanisms underlying boundary formation are not well understood. In this study, we focused on S. cerevisiae ADA1, a gene previously shown to encode a protein with a robust boundary function. Ada1 is a component of the histone modification complex Spt-Ada-Gcn5-acetyltransferase (SAGA) and the SAGA-like (SLIK) complex, and it helps to maintain the integrity of these complexes...
October 2016: Genes to Cells: Devoted to Molecular & Cellular Mechanisms
https://www.readbyqxmd.com/read/27642009/the-saga-complex-in-the-rice-pathogen-fusarium-fujikuroi-structure-and-functional-characterization
#19
Sarah M Rösler, Katharina Kramer, Iris Finkemeier, Hans-Ulrich Humpf, Bettina Tudzynski
Post-translational modification of histones is a crucial mode of transcriptional regulation in eukaryotes. A well-described acetylation modifier of certain lysine residues is the Spt-Ada-Gcn5 acetyltransferase (SAGA) complex assembled around the histone acetyltransferase Gcn5 in Saccharomyces cerevisiae. We identified and characterized the SAGA complex in the rice pathogen Fusarium fujikuroi, well-known for producing a large variety of secondary metabolites (SMs). By using a co-immunoprecipitation approach, almost all of the S...
September 19, 2016: Molecular Microbiology
https://www.readbyqxmd.com/read/27601672/structural-mechanism-for-the-recognition-and-ubiquitination-of-a-single-nucleosome-residue-by-rad6-bre1
#20
Laura D Gallego, Medini Ghodgaonkar Steger, Anton A Polyansky, Tobias Schubert, Bojan Zagrovic, Ning Zheng, Tim Clausen, Franz Herzog, Alwin Köhler
Cotranscriptional ubiquitination of histone H2B is key to gene regulation. The yeast E3 ubiquitin ligase Bre1 (human RNF20/40) pairs with the E2 ubiquitin conjugating enzyme Rad6 to monoubiquitinate H2B at Lys123. How this single lysine residue on the nucleosome core particle (NCP) is targeted by the Rad6-Bre1 machinery is unknown. Using chemical cross-linking and mass spectrometry, we identified the functional interfaces of Rad6, Bre1, and NCPs in a defined in vitro system. The Bre1 RING domain cross-links exclusively with distinct regions of histone H2B and H2A, indicating a spatial alignment of Bre1 with the NCP acidic patch...
September 20, 2016: Proceedings of the National Academy of Sciences of the United States of America
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