A Germline Clone Screen for Meiotic Mutants in<i>Drosophila melanogaster</i>

Page, Scott L.; Nielsen, Rachel J.; Teeter, Kathy; Lake, Cathleen M.; Ong, Seng-Kai; Wright, Kathleen R.; Dean, Kristen L.; Agne, Daniel; Gilliland, William D.; Hawley, R. Scott

doi:10.4161/fly.4720

Cited by 26 publications

(50 citation statements)

References 65 publications

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“…w ; pCa4-attB-genomic-Corolla (this study), okra AA cn bw / CyO (Ghabrial et al 1998), okra RU cn bw /CyO (Ghabrial et al 1998), ru 1 h 1 th 1 st 1 cu 1 sr 1 e s ca 1 (Bloomington 576), ru 1 h 1 th 1 st 1 cu 1 sr 1 e s Pr 1 ca / TM6B , Bri 1 Tb 1 (Bloomington 1711), and ru 1 h 1 th 1 st 1 cu 1 sr 1 e s ca 1 / TM3, Sb were used in recombination assays. Other stocks used: nanos-Gal4 ; cona f04903 / TM3 , y w eyFLP ; FRT82B y + TPN1 , TbSer (Page et al 2008), yw eyFLP;FRT82B cona A12 / TM6B y + TPN1 , Tb (Page et al 2007), y w FRT19A / hs-hidY (used as wild-type stock) (Collins et al 2012), y w nanos-Gal4 / FM7a ; c(3)G 68 e ca / TM3, Ser (Jeffress et al 2007), y w ; P{UASP}-c(3)G Cdel4-Flag ; c(3)G [68] e / TM3 , Sb Ser e ; spa pol /+ (Jeffress et al 2007), and w / B s Y ; c(3)G 68 e / TM3 , Ser .…”

Section: Methodsmentioning

confidence: 99%

Corolla Is a Novel Protein That Contributes to the Architecture of the Synaptonemal Complex of Drosophila

et al. 2014

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In most organisms the synaptonemal complex (SC) connects paired homologs along their entire length during much of meiotic prophase. To better understand the structure of the SC, we aim to identify its components and to determine how each of these components contributes to SC function. Here, we report the identification of a novel SC component in Drosophila melanogaster female oocytes, which we have named Corolla. Using structured illumination microscopy, we demonstrate that Corolla is a component of the central region of the SC. Consistent with its localization, we show by yeast two-hybrid analysis that Corolla strongly interacts with Cona, a central element protein, demonstrating the first direct interaction between two inner-synaptonemal complex proteins in Drosophila. These observations help provide a more complete model of SC structure and function in Drosophila females.

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Section: Methodsmentioning

confidence: 99%

Corolla Is a Novel Protein That Contributes to the Architecture of the Synaptonemal Complex of Drosophila

et al. 2014

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“…The wild-type strain used in this study was yw eyFLP; FRT82, which was the strain used to induce the EMS mutation (Page et al 2007). Deficiency strains used for mapping were obtained from the Bloomington Drosophila Stock Center.…”

Section: Methodsmentioning

confidence: 99%

“…Overview of the isolation of an EMS-induced mutant in mcm5: A genetic screen was designed to recover new meiotic mutants that did not exclude essential genes (details of screen in Page et al 2007). Briefly, we adapted the FLP-FRT system to design a screen that required a meiotic nondisjunction event of an autosome to survive the screen.…”

Section: Methodsmentioning

confidence: 99%

“…Mapping and identification of an allele of mcm5: The EMSinduced lesion in A7 was mapped using indel and deficiency mapping (for details see Page et al 2007). Briefly, recombinants are made between the unmarked FRT-bearing chromosome containing A7 and a chromosome containing a distal marker element known as EP.…”

Section: Methodsmentioning

confidence: 99%

“…This screen is described in detail in Page et al (2007), but briefly the right arms of third chromosomes mutagenized with EMS were made homozygous in the female germline using FLP/ FRT-mediated recombination. Females carrying such germline clones were then mated to appropriate tester males to screen for those mutants that caused high levels of autosomal nondisjunction.…”

Section: A7mentioning

confidence: 99%

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A Genetic Analysis of the Drosophilamcm5Gene Defines a Domain Specifically Required for Meiotic Recombination

et al. 2007

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Members of the minichromosome maintenance (MCM) family have pivotal roles in many biological processes. Although originally studied for their role in DNA replication, it is becoming increasingly apparent that certain members of this family are multifunctional and also play roles in transcription, cohesion, condensation, and recombination. Here we provide a genetic dissection of the mcm5 gene in Drosophila that demonstrates an unexpected function for this protein. First, we show that homozygotes for a null allele of mcm5 die as third instar larvae, apparently as a result of blocking those replication events that lead to mitotic divisions without impairing endo-reduplication. However, we have also recovered a viable and fertile allele of mcm5 (denoted mcm5 A7) that specifically impairs the meiotic recombination process. We demonstrate that the decrease in recombination observed in females homozygous for mcm5 A7 is not due to a failure to create or repair meiotically induced double strand breaks (DSBs), but rather to a failure to resolve those DSBs into meiotic crossovers. Consistent with their ability to repair meiotically induced DSBs, flies homozygous for mcm5 A7 are fully proficient in somatic DNA repair. These results strengthen the observation that members of the prereplicative complex have multiple functions and provide evidence that mcm5 plays a critical role in the meiotic recombination pathway.

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Synaptonemal Complex-Dependent Centromeric Clustering and the Initiation of Synapsis in Drosophila Oocytes

et al. 2011

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The pairing of homologous chromosomes and the intimate synapsis of the paired homologs by the synaptonemal complex (SC) are essential for subsequent meiotic processes including recombination and chromosome segregation. Here we show that the centromere clustering plays an important role in initiating homolog synapsis during meiosis in Drosophila females. Although centromeres are not clustered prior to the onset of meiosis, all four pairs of centromeres are actively clustered into one or two masses during early meiotic prophase. Within the 16-cell cyst, centromeric clustering appears to define the first step in the initiation of synapsis. Clustering is restricted to the nuclei that form the SC and is dependent on all known SC proteins. Surprisingly, both centromeric clusters and the SC components associated with them persist long after the disassembly of the euchromatic SC at the end of pachytene. The initiation of homologous recombination through the formation of programmed double-strand breaks (DSBs) is not required for either the formation or the maintenance of the centromeric clusters. Our data support a view in which the SC-mediated clustering at the centromeres is the initiating event for meiotic synapsis.

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A Germline Clone Screen for Meiotic Mutants inDrosophila melanogaster

Cited by 26 publications

References 65 publications

Corolla Is a Novel Protein That Contributes to the Architecture of the Synaptonemal Complex of Drosophila

Corolla Is a Novel Protein That Contributes to the Architecture of the Synaptonemal Complex of Drosophila

A Genetic Analysis of the Drosophilamcm5Gene Defines a Domain Specifically Required for Meiotic Recombination

Synaptonemal Complex-Dependent Centromeric Clustering and the Initiation of Synapsis in Drosophila Oocytes

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