Browsing by Author "Burnetti, Anthony J"
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Item Open Access Cell cycle Start is coupled to entry into the yeast metabolic cycle across diverse strains and growth rates.(Mol Biol Cell, 2016-01-01) Burnetti, Anthony J; Aydin, Mert; Buchler, Nicolas ECells have evolved oscillators with different frequencies to coordinate periodic processes. Here we studied the interaction of two oscillators, the cell division cycle (CDC) and the yeast metabolic cycle (YMC), in budding yeast. Previous work suggested that the CDC and YMC interact to separate high oxygen consumption (HOC) from DNA replication to prevent genetic damage. To test this hypothesis, we grew diverse strains in chemostat and measured DNA replication and oxygen consumption with high temporal resolution at different growth rates. Our data showed that HOC is not strictly separated from DNA replication; rather, cell cycle Start is coupled with the initiation of HOC and catabolism of storage carbohydrates. The logic of this YMC-CDC coupling may be to ensure that DNA replication and cell division occur only when sufficient cellular energy reserves have accumulated. Our results also uncovered a quantitative relationship between CDC period and YMC period across different strains. More generally, our approach shows how studies in genetically diverse strains efficiently identify robust phenotypes and steer the experimentalist away from strain-specific idiosyncrasies.Item Open Access Coupling of the Yeast Metabolic Cycle and the Cell Division Cycle in Populations and Single Cells(2017) Burnetti, Anthony JBiological oscillators are ubiquitous in living systems. They allow cellular processes to anticipate and act in synchrony with regular events in the outside world (such as the day/night cycle), or they ensure that processes occur in a particular order. Living things typically contain multiple oscillators, which can often couple to each other and influence each other's timing and function. The purpose of this thesis has been to investigate the relationship between two coupled oscillators in \textit{Saccharomyces cerevisiae}: the yeast metabolic cycle and the cell division cycle. I have focused on two key questions: what is the biological significance of their coupling, and is one oscillator dominant in its interaction with the other?
First, I investigated the temporal relationship between the cell division cycle and metabolic shifts that occur during the metabolic cycle across diverse yeast strains. I showed that a particular cell cycle event (DNA replication) was consistently delayed relative to a metabolic event (entry into the high oxygen consumption phase). This suggested that an earlier cell cycle event (Start and commitment to the cell cycle) was tied to the onset of high oxygen consumption. Second, I used fluorescent probes to examine the relationship between the metabolic cycle and the commitment to cell cycle progression at single-cell resolution. This revealed that cells enter high oxygen consumption phase of the metabolic cycle before passing Start, supporting a model of metabolic cycle/cell division cycle coupling in which the shorter metabolic cycle controls cell cycle commitment, likely via modulation of cell size thresholds.
Item Open Access The exon junction complex component Magoh controls brain size by regulating neural stem cell division.(Nat Neurosci, 2010-05) Silver, Debra L; Watkins-Chow, Dawn E; Schreck, Karisa C; Pierfelice, Tarran J; Larson, Denise M; Burnetti, Anthony J; Liaw, Hung-Jiun; Myung, Kyungjae; Walsh, Christopher A; Gaiano, Nicholas; Pavan, William JBrain structure and size require precise division of neural stem cells (NSCs), which self-renew and generate intermediate neural progenitors (INPs) and neurons. The factors that regulate NSCs remain poorly understood, and mechanistic explanations of how aberrant NSC division causes the reduced brain size seen in microcephaly are lacking. Here we show that Magoh, a component of the exon junction complex (EJC) that binds RNA, controls mouse cerebral cortical size by regulating NSC division. Magoh haploinsufficiency causes microcephaly because of INP depletion and neuronal apoptosis. Defective mitosis underlies these phenotypes, as depletion of EJC components disrupts mitotic spindle orientation and integrity, chromosome number and genomic stability. In utero rescue experiments showed that a key function of Magoh is to control levels of the microcephaly-associated protein Lis1 during neurogenesis. Our results uncover requirements for the EJC in brain development, NSC maintenance and mitosis, thereby implicating this complex in the pathogenesis of microcephaly.