A New Family of Drosophila Balancer Chromosomes With a <i>w− dfd</i>-GMR Yellow Fluorescent Protein Marker

Le, Tien; Liang, Zhiting; Patel, Heeren; Yu, Marcus; Sivasubramaniam, Gitanjali; Slovitt, Matthew; Tanentzapf, Guy; Mohanty, Nihar Nalini; Paul, Sarah M.; Wu, Victoria; Beitel, Greg J.

doi:10.1534/genetics.106.063461

Cited by 86 publications

(72 citation statements)

References 14 publications

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“…The UAS-kune-RNAi 108224 line was used for all RNAi experiments. The FM7 dfd-YFP, CyO dfd-YFP, CyO act-Z, TM6 dfd-YFP, and TM6 dfd-Z balancers were used for genotyping single and double mutants (Le et al 2006). mega; kune; sinu triple mutants were identified by the absence of all three proteins, as assessed by immunohistochemistry.…”

Section: Methodsmentioning

confidence: 99%

The Drosophila Claudin Kune-kune Is Required for Septate Junction Organization and Tracheal Tube Size Control

2010

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The vertebrate tight junction is a critical claudin-based cell-cell junction that functions to prevent free paracellular diffusion between epithelial cells. In Drosophila, this barrier is provided by the septate junction, which, despite being ultrastructurally distinct from the vertebrate tight junction, also contains the claudin-family proteins Megatrachea and Sinuous. Here we identify a third Drosophila claudin, Kunekune, that localizes to septate junctions and is required for junction organization and paracellular barrier function, but not for apical-basal polarity. In the tracheal system, septate junctions have a barrierindependent function that promotes lumenal secretion of Vermiform and Serpentine, extracellular matrix modifier proteins that are required to restrict tube length. As with Sinuous and Megatrachea, loss of Kune-kune prevents this secretion and results in overly elongated tubes. Embryos lacking all three characterized claudins have tracheal phenotypes similar to any single mutant, indicating that these claudins act in the same pathway controlling tracheal tube length. However, we find that there are distinct requirements for these claudins in epithelial septate junction formation. Megatrachea is predominantly required for correct localization of septate junction components, while Sinuous is predominantly required for maintaining normal levels of septate junction proteins. Kune-kune is required for both localization and levels. Double-and triple-mutant combinations of Sinuous and Megatrachea with Kunekune resemble the Kune-kune single mutant, suggesting that Kune-kune has a more central role in septate junction formation than either Sinuous or Megatrachea.

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Section: Methodsmentioning

confidence: 99%

The Drosophila Claudin Kune-kune Is Required for Septate Junction Organization and Tracheal Tube Size Control

2010

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“…LL02791 (Schuldiner et al 2008) (Le et al 2006) were used for the identification of double mutant embryos and larvae based on absence of EYFP fluorescence in the head region.…”

Section: Pbac{sastopdsred}mentioning

confidence: 99%

Drosophila Nnf1 paralogs are partially redundant for somatic and germ line kinetochore function

et al. 2016

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Kinetochores allow attachment of chromosomes to spindle microtubules. Moreover, they host proteins that permit correction of erroneous attachments and prevent premature anaphase onset before bi-orientation of all chromosomes in metaphase has been achieved. Kinetochores are assembled from subcomplexes. Kinetochore proteins as well as the underlying centromere proteins and the centromeric DNA sequences evolve rapidly despite their fundamental importance for faithful chromosome segregation during mitotic and meiotic divisions. During evolution of Drosophila melanogaster, several centromere proteins were lost and a recent gene duplication has resulted in two Nnf1 paralogs, Nnf1a and Nnf1b, which code for alternative forms of a Mis12 kinetochore complex component. The rapid evolutionary divergence of centromere/kinetochore constituents in animals and plants has been proposed to be driven by an intragenome conflict resulting from centromere drive during female meiosis. Thus, a female meiosisspecific paralog might be expected to evolve rapidly under positive selection. While our characterization of the D. melanogaster Nnf1 paralogs hints at some partial functional specialization of Nnf1b for meiosis, we have failed to detect evidence for positive selection in our analysis of Nnf1 sequence evolution in the Drosophilid lineage. Neither paralog is essential, even though we find some clear differences in subcellular localization and expression during development. Loss of both paralogs results in developmental lethality. We therefore conclude that the two paralogs are still in early stages of differentiation. Neither paralog is essential, even though we find some clear differences in subcellular localization and expression during development. Loss of both paralogs results in developmental lethality. We therefore conclude that the two paralogs are still in early stages of differentiation.

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“…The breathless promoter btl-Gal4 was used in combination with UAS-GFP as the reporter in the X-chromosome complementation tests (Shiga et al 1996). For the balancer chromosomes, we used w*; ry 506 Dr 1 /TM6B, P{Dfd-EYFP} Sb 1 ca 1 (Le et al 2006) and FM7i, P{ActGFP}JMR3 (Reichhart and Ferrandon 1998). Complementation tests and deficiency experiments for Smg6 were performed using Df(3R)ED6220/TM6c Tb, Sb (Ryder et al 2007).…”

Section: Apmentioning

confidence: 99%

Drosophila mutants show NMD pathway activity is reduced, but not eliminated, in the absence of Smg6

Frizzell¹,

Rynearson²,

Metzstein³

2012

RNA

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The nonsense-mediated mRNA decay (NMD) pathway is best known for targeting mutant mRNAs containing premature termination codons for rapid degradation, but it is also required for regulation of many endogenous transcripts. Components of the NMD pathway were originally identified by forward genetic screens in yeast and Caenorhabditis elegans. In other organisms, the NMD pathway has been investigated by studying the homologs of these genes. We present here the first unbiased genetic screen in Drosophila designed specifically to identify genes involved in NMD. By using a highly efficient genetic mosaic approach, we have screened~40% of the Drosophila genome and isolated more than 40 alleles of genes required for NMD. We focus on alleles we have obtained in two known NMD components: Upf2 and Smg6. Our analysis of multiple alleles of the core NMD component Upf2 reveals that the Upf2 requirement in NMD may be separate from its requirement for viability, indicating additional critical cellular roles for this protein. Our alleles of Smg6 are the first point mutations obtained in Drosophila, and we find that Smg6 has both endonucleolytic and nonendonucleolytic roles in NMD. Thus, our genetic screens have revealed that Drosophila NMD factors play distinct roles in target regulation, similar to what is found in mammals, but distinct from the relatively similar requirements for NMD genes observed in C. elegans and yeast.

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A New Family of Drosophila Balancer Chromosomes With a w− dfd-GMR Yellow Fluorescent Protein Marker

Cited by 86 publications

References 14 publications

The Drosophila Claudin Kune-kune Is Required for Septate Junction Organization and Tracheal Tube Size Control

The Drosophila Claudin Kune-kune Is Required for Septate Junction Organization and Tracheal Tube Size Control

Drosophila Nnf1 paralogs are partially redundant for somatic and germ line kinetochore function

Drosophila mutants show NMD pathway activity is reduced, but not eliminated, in the absence of Smg6

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