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1.
PLoS Comput Biol ; 12(3): e1004825, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27010473

RESUMO

Metabolic efficiency depends on the balance between supply and demand of metabolites, which is sensitive to environmental and physiological fluctuations, or noise, causing shortages or surpluses in the metabolic pipeline. How cells can reliably optimize biomass production in the presence of metabolic fluctuations is a fundamental question that has not been fully answered. Here we use mathematical models to predict that enzyme saturation creates distinct regimes of cellular growth, including a phase of growth arrest resulting from toxicity of the metabolic process. Noise can drive entry of single cells into growth arrest while a fast-growing majority sustains the population. We confirmed these predictions by measuring the growth dynamics of Escherichia coli utilizing lactose as a sole carbon source. The predicted heterogeneous growth emerged at high lactose concentrations, and was associated with cell death and production of antibiotic-tolerant persister cells. These results suggest how metabolic networks may balance costs and benefits, with important implications for drug tolerance.


Assuntos
Pontos de Checagem do Ciclo Celular/fisiologia , Enzimas/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/citologia , Escherichia coli/crescimento & desenvolvimento , Modelos Biológicos , Proliferação de Células/fisiologia , Simulação por Computador , Ativação Enzimática , Taxa de Depuração Metabólica , Proteínas de Transporte de Monossacarídeos/metabolismo , Simportadores/metabolismo , beta-Galactosidase/metabolismo
2.
PLoS Genet ; 11(8): e1005332, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26241928

RESUMO

Sexual reproduction allows transposable elements (TEs) to proliferate, leading to rapid divergence between populations and species. A significant outcome of divergence in the TE landscape is evident in hybrid dysgenic syndromes, a strong form of genomic incompatibility that can arise when (TE) family abundance differs between two parents. When TEs inherited from the father are absent in the mother's genome, TEs can become activated in the progeny, causing germline damage and sterility. Studies in Drosophila indicate that dysgenesis can occur when TEs inherited paternally are not matched with a pool of corresponding TE silencing PIWI-interacting RNAs (piRNAs) provisioned by the female germline. Using the D. virilis syndrome of hybrid dysgenesis as a model, we characterize the effects that divergence in TE profile between parents has on offspring. Overall, we show that divergence in the TE landscape is associated with persisting differences in germline TE expression when comparing genetically identical females of reciprocal crosses and these differences are transmitted to the next generation. Moreover, chronic and persisting TE expression coincides with increased levels of genic piRNAs associated with reduced gene expression. Combined with these effects, we further demonstrate that gene expression is idiosyncratically influenced by differences in the genic piRNA profile of the parents that arise though polymorphic TE insertions. Overall, these results support a model in which early germline events in dysgenesis establish a chronic, stable state of both TE and gene expression in the germline that is maintained through adulthood and transmitted to the next generation. This work demonstrates that divergence in the TE profile is associated with diverse piRNA-mediated transgenerational effects on gene expression within populations.


Assuntos
Elementos de DNA Transponíveis , Drosophila/genética , RNA Interferente Pequeno/genética , Alelos , Animais , Quimera/genética , Drosophila/metabolismo , Epigênese Genética , Feminino , Expressão Gênica , Genes de Insetos , Masculino , Ovário/metabolismo , RNA Interferente Pequeno/metabolismo
3.
Biotechniques ; 55(5): 269-72, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24215643

RESUMO

A large number of methods are available to deplete ribosomal RNA reads from high-throughput RNA sequencing experiments. Such methods are critical for sequencing Drosophila small RNAs between 20 and 30 nucleotides because size selection is not typically sufficient to exclude the highly abundant class of 30 nucleotide 2S rRNA. Here we demonstrate that pre-annealing terminator oligos complimentary to Drosophila 2S rRNA prior to 5' adapter ligation and reverse transcription efficiently depletes 2S rRNA sequences from the sequencing reaction in a simple and inexpensive way. This depletion is highly specific and is achieved with minimal perturbation of miRNA and piRNA profiles.


Assuntos
Drosophila/genética , RNA Ribossômico/genética , Pequeno RNA não Traduzido/genética , Análise de Sequência de RNA/métodos , Animais , Sequência de Bases , MicroRNAs/genética , Dados de Sequência Molecular
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