Cangaia de Jegue and DamID

Contents 1 Career 2 Members 3 Discography 3.1 Albums 3.2 Singles 4 References 5 External links

Career

In August 2002, four Brazilian university students got together in Jequié nicknamed "A cidade do sol" (city of sun) forming a band that played at university gigs and various social occasions. They wanted to call it the Cangaia band (local slang for "yoke" put on animals to carry the load). But discovering that the name was already used by another band, they decided to add "of something" so they picked the donkey (jegue) renaming the band "Cangaia de Jegue" (the yoke of the donkey) in a cheeky reference to the commonly known area "República dos Jegues" (the republic of donkeys).

Becoming instantly popular in their city, singer Norberto Curvello penned the song "Saudade do Interior" in gratitude followed by the 2004 self-titled album Cangaia de Jegue, with many tracks becoming hits on local Bahia radio, most notably the songs "O beijo teu" (Your Kiss) and "De frente pro mar" (Beachfront) with the latter becoming popular with Estakazero, a well-established local band, doing a cover of the Cangaia track. Cangaia also took part in João de Jequié, Forró da Margarida, Forró da Onça, and were regular features at Jaguaquara Coffe and Forró Coffe, in Itiruçu.

In 2006 the band moved to Bahia's bigger coastline city Salvador for bigger opportunities, releasing their second album Você vai ver (meaning You will see) following it up with a DVD launched in Jequié first and then in other cities, thus competing with better known bands like Estakazero and Seu Maxixe.

After the success of the song "Ai se eu te pego!" by Michel Teló, Cangaia de Jegue are enjoying a huge increase in their popularity throughout Brazil as fans check the earlier version of the song before Michel Teló's release. Members

The initial formation of the band was just 4 members Norberto Curvêllo (vocals and guitar) Marcelo Capucho (bass drums) Junior Bomfim (triangle) Humberto Júnior (bass)

The present day formation is made up of 6 permanent members: Norberto Curvêllo (vocals and guitar) Junior Bomfim (triangle and vocals) Allê Barbosa (bass) Clécio Carvalho (accordion) Serginho di Boca (bass drums, drums, vocals). Bruno Valverde (guitar).

Sometimes Marcelo Tribal also takes part with bass drum and percussion. Discography Albums 2004: Cangaia de Jegue 2006: Você vai ver (The album contains collaboration with Leo Macedo (of band Estakazero)) 2009: Cangaia Inté o Sol Raia (CD / DVD) (Contains collaborations with Circuladô de fulo, the trio Virgulino, Rastapé and Waldo) 2009: Cangaia de Jegue – Ao Vivo) (live album) 2010: Ai se eu te pego (promotional CD)

Tracklist "Chama essa cerveja" "Dedinho na boca" "Cachaça véa" "Ai se eu te pego" "Solteiro de novo" "Sei não" "Balada" "Cachaça na mão" "Mulher danada" "Do jeito que tu vaz e carinhoso" Coração tapado" "Agora é beber" "Galera louca" "Neide" "Festa de camisa" "Triste, allegre" "Me chama de my love" "Sinto falta" (bonus) Singles 2004: "Saudade do Interior" 2006: "O beijo teu" 2006: "De frente pro mar" 2008: "Ai se eu te pego" 2009: "Dança do Ice" 2011: "Bolo doido"

DamID and Cangaia de Jegue

Contents 1 Description of the method 1.1 Principle 1.2 Methyl PCR (mePCR) 1.3 Specificities of DamID versus ChIP (Chromatin Immuno-Precipitation) 2 Known biases and technical issues 2.1 Plasmid methylation bias 2.2 Apoptosis 2.3 Resolution 3 References 4 External links

Description of the method Principle Principle of DamID. This sketch shows an idealized view of the DNA molecule wrapped around histones within the nucleus of a cell. The enzyme Dam (green) is fused to the protein of interest (orange) by expression of a chimeric DNA sequence. The protein of interest drags Dam onto its cognate targets. The tethering leads to methylation of GATCs in the neighborhood of the binding site (red) but not at a distance.

N6-methyladenine (m6A) is the product of the addition of a methyl group (CH3) at position 6 of the adenine. This modified nucleotide is absent from the vast majority of eukaryotes, but is widespread in bacterial genomes, as part of the restriction modification or DNA repair systems. In Escherichia coli, adenine methylation is catalyzed by the adenine methyltransferase Dam (DNA adenine methyltransferase), which catalyses adenine methylation exclusively in the palindromic sequence GATC. Ectopic expression of Dam in eukaryotic cells leads to methylation of adenine in GATC sequences without any other noticeable side effect.

Based on this, DamID consists in fusing Dam to a protein of interest (usually a protein that interacts with DNA such as transcription factors) or a chromatin component. The protein of interest thus targets Dam to its cognate in vivo binding site, resulting in the methylation of neighboring GATCs. The presence of m6A, coinciding with the binding sites of the proteins of interest, is revealed by methyl PCR. Methyl PCR (mePCR)

In this assay the genome is digested by DpnI, which cuts only methylated GATCs. Double-stranded adapters with a known sequence are then ligated to the ends generated by DpnI. A PCR with primers matching the adaptors is then carried out, leading to the specific amplification of genomic fragments flanked by methylated GATCs. In practice, ligation products are digested by DpnII prior PCR amplification. This enzyme cuts non-methylated GATCs, ensuring that only fragments flanked by consecutive methylated GATCs are amplified. Specificities of DamID versus ChIP (Chromatin Immuno-Precipitation)

ChIP is an alternative method to assay protein binding at specific loci of the genome. Unlike ChIP, DamID does not require a specific antibody against the protein of interest. On the one hand, this allows to map proteins for which no such antibody is available. On the other hand, this makes it impossible to specifically map posttranslationally modified proteins.

Another fundamental difference is that ChIP assays where the protein of interests is at a given time, whereas DamID assays where it has been. The reason is that m6A stays in the DNA after the Dam fusion protein goes away. For proteins that are either bound or unbound on their target sites this does not change the big picture. However, this can lead to strong differences in the case of proteins that slide along the DNA (e.g. RNA polymerase). Known biases and technical issues Plasmid methylation bias

Depending on how the experiment is carried out, DamID can be subject to plasmid methylation biases. Because plasmids are usually amplified in E. coli where Dam is naturally expressed, they are methylated on every GATC. In transient transfection experiments, the DNA of those plasmids is recovered along with the DNA of the transfected cells, meaning that fragments of the plasmid are amplified in the mePCR. Every sequence of the genome that shares homology or identity with the plasmid may thus appear to be bound by the protein of interest. In particular, this is true of the open reading frame of the protein of interest, which is present in both the plasmid and the genome. In microarray experiments, this bias can be used to ensure that the proper material was hybridized. Apoptosis

Apoptotic cells degrade their DNA in a characteristic nucleosome ladder pattern. This generates DNA fragments that can be ligated and amplified during the DamID procedure (van Steensel laboratory, unpublished observations). The influence of these nucleosomal fragments on the binding profile of a protein is not known. Resolution

The resolution of DamID is a function of the availability of GATC sequences in the genome. A protein can only be mapped within two consecutive GATC sites. The median spacing between GATC fragments is 205 bp in Drosophila (FlyBase release 5), 260 in mouse (Mm9), and 460 in human (HG19).
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