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1.
J Cell Sci ; 131(9)2018 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-29643118

RESUMO

In Drosophila, zygotic genome activation occurs in pre-blastoderm embryos during rapid mitotic divisions. How the transcription machinery is coordinated to achieve this goal in a very brief time span is still poorly understood. Transcription factor II H (TFIIH) is fundamental for transcription initiation by RNA polymerase II (RNAPII). Herein, we show the in vivo dynamics of TFIIH at the onset of transcription in Drosophila embryos. TFIIH shows an oscillatory behaviour between the nucleus and cytoplasm. TFIIH foci are observed from interphase to metaphase, and colocalize with those for RNAPII phosphorylated at serine 5 (RNAPIIS5P) at prophase, suggesting that transcription occurs during the first mitotic phases. Furthermore, embryos with defects in subunits of either the CAK or the core subcomplexes of TFIIH show catastrophic mitosis. Although, transcriptome analyses show altered expression of several maternal genes that participate in mitosis, the global level of RNAPIIS5P in TFIIH mutant embryos is similar to that in the wild type, therefore, a direct role for TFIIH in mitosis cannot be ruled out. These results provide important insights regarding the role of a basal transcription machinery component when the zygotic genome is activated.


Assuntos
Drosophila/embriologia , Drosophila/genética , Genômica/métodos , Fator de Transcrição TFIIH/metabolismo , Animais , Animais Geneticamente Modificados , Linhagem Celular , Drosophila/metabolismo , Feminino , Masculino , Mitose/fisiologia , Zigoto
2.
J Cell Biochem ; 120(3): 3887-3897, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30270456

RESUMO

Alternatives to the cap mechanism in translation are often used by viruses and cells to allow them to synthesize proteins in events of stress and viral infection. In Drosophila there are hundreds of polycistronic messenger RNA (mRNA), and various mechanisms are known to achieve this. However, proteins in a same mRNA often work in the same cellular mechanism, this is not the case for Drosophila's Swc6/p18Hamlet homolog Dmp18, part of the SWR1 chromatin remodeling complex, who is encoded in a bicistronic mRNA next to Dmp8 (Dmp8-Dmp18 transcript), a structural component of transcription factor TFIIH. The organization of these two genes as a bicistron is conserved in all arthropods, however the length of the intercistronic sequence varies from more than 90 to 2 bases, suggesting an unusual translation mechanism for the second open reading frame. We found that even though translation of Dmp18 occurs independently from that of Dmp8, it is necessary for Dmp18 to be in that conformation to allow its correct translation during cellular stress caused by damage via heat-shock and UV radiation.


Assuntos
Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Biossíntese de Proteínas , RNA Mensageiro/genética , Fator de Transcrição TFIIH/genética , Sequência de Aminoácidos , Animais , Cromatina/química , Cromatina/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Regulação da Expressão Gênica , Temperatura Alta , Larva/genética , Larva/metabolismo , Fases de Leitura Aberta , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Estresse Fisiológico , Fator de Transcrição TFIIH/metabolismo , Raios Ultravioleta
3.
J Biol Chem ; 285(41): 31370-9, 2010 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-20675387

RESUMO

Chromatin undergoes a variety of changes in response to UV-induced DNA damage, including histone acetylation. In human and Drosophila cells, this response is affected by mutations in the tumor suppressor p53. In this work, we report that there is a global decrease in trimethylated Lys-9 in histone H3 (H3K9me3) in salivary gland cells in wild type flies in response to UV irradiation. In contrast, flies with mutations in the Dmp53 gene have reduced basal levels of H3K9me3, which are then increased after UV irradiation. The reduction of H3K9me3 in response to DNA damage occurs preferentially in heterochromatin. Our experiments demonstrate that UV irradiation enhances the levels of Lys-9 demethylase (dKDM4B) transcript and protein in wild type flies, but not in Dmp53 mutant flies. Dmp53 binds to a DNA element in the dKdm4B gene as a response to UV irradiation. Furthermore, heterozygous mutants for the dKdm4B gene are more sensitive to UV irradiation; they are deficient in the removal of cyclobutane-pyrimidine dimers, and the decrease of H3K9me3 levels following DNA damage is not observed in dKdm4B mutant flies. We propose that in response to UV irradiation, Dmp53 enhances the expression of the dKDM4B histone demethylase, which demethylates H3K9me3 preferentially in heterochromatin regions. This mechanism appears to be essential for the proper function of the nucleotide excision repair system.


Assuntos
Dano ao DNA/efeitos da radiação , Proteínas de Drosophila/metabolismo , Heterocromatina/metabolismo , Histona Desmetilases/metabolismo , Histonas/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Raios Ultravioleta/efeitos adversos , Animais , Dano ao DNA/genética , Reparo do DNA/genética , Reparo do DNA/efeitos da radiação , Proteínas de Drosophila/genética , Drosophila melanogaster , Heterocromatina/genética , Histona Desmetilases/genética , Histonas/genética , Humanos , Lisina/genética , Lisina/metabolismo , Mutação , Dímeros de Pirimidina/genética , Dímeros de Pirimidina/metabolismo , Proteína Supressora de Tumor p53/genética
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