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Julie M Claycomb

Publications and source records attributed to Julie M Claycomb.

3 recordsLinked to original sources

Developmental gene amplification: insights into DNA replication and gene expression.

In the formation of a complex organism and the differentiation of specific cell types, there are often demands for high levels of particular gene products. These demands can be met by increasing transcription or translation, or by decreasing the rate of mRNA or protein turnover. Although these are the most common means to increase expression levels, there is another mechanism: gene amplification. Developmental gene amplification is a DNA replication-based process whereby specific genes are replicated above the copy number of surrounding sequences, resulting in an increase in the template available for transcription. Recent microarray studies in Drosophila melanogaster have identified two additional amplicons, suggesting that developmental gene amplification might be more widely used than was previously thought. Furthermore, work in both Drosophila and the related fly, Sciara coprophila, has yielded insights into the mechanisms, regulatory sequences and proteins controlling DNA replication during gene amplification, including a link between transcription factors and origin usage.

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Gene amplification as a developmental strategy: isolation of two developmental amplicons in Drosophila.

Gene amplification is known to be critical for upregulating gene expression in a few cases, but the extent to which amplification is utilized in the development of diverse organisms remains unknown. By quantifying genomic DNA hybridization to microarrays to assay gene copy number, we identified two additional developmental amplicons in the follicle cells of the Drosophila ovary. Both amplicons contain genes which, following their amplification, are expressed in the follicle cells, and the expression of three of these genes becomes restricted to specialized follicle cells late in differentiation. Genetic analysis establishes that at least one of these genes, yellow-g, is critical for follicle cell function, because mutations in yellow-g disrupt eggshell integrity. Thus, during follicle cell differentiation the entire genome is overreplicated as the cells become polyploid, and subsequently specific genomic intervals are overreplicated to facilitate gene expression.

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Visualization of replication initiation and elongation in Drosophila.

Chorion gene amplification in the ovaries of Drosophila melanogaster is a powerful system for the study of metazoan DNA replication in vivo. Using a combination of high-resolution confocal and deconvolution microscopy and quantitative realtime PCR, we found that initiation and elongation occur during separate developmental stages, thus permitting analysis of these two phases of replication in vivo. Bromodeoxyuridine, origin recognition complex, and the elongation factors minichromosome maintenance proteins (MCM)2-7 and proliferating cell nuclear antigen were precisely localized, and the DNA copy number along the third chromosome chorion amplicon was quantified during multiple developmental stages. These studies revealed that initiation takes place during stages 10B and 11 of egg chamber development, whereas only elongation of existing replication forks occurs during egg chamber stages 12 and 13. The ability to distinguish initiation from elongation makes this an outstanding model to decipher the roles of various replication factors during metazoan DNA replication. We utilized this system to demonstrate that the pre-replication complex component, double-parked protein/cell division cycle 10-dependent transcript 1, is not only necessary for proper MCM2-7 localization, but, unexpectedly, is present during elongation.

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