The urate oxidase gene of Drosophila pseudoobscura and Drosophila melanogaster: Evolutionary changes of sequence and regulation

Tb, Friedman; Jb, Burnett; Lootens, S; Steinman, Richard A.; Wallrath, Lori L.

doi:10.1007/bf00163853

Cited by 12 publications

(5 citation statements)

References 103 publications

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“…To determine the expression levels of Drosophila Lamin C, proteins were extracted from third instar larvae (78) and separated by size on 10–12% polyacrylamide gels, transferred to nitrocellulose membrane and incubated with anti-Lamin C LC28.26 anti-mouse IgG (11) used at 1:5000–1:8000 dilution or anti-alpha tubulin anti-mouse IgG1 [Sigma (St. Louis) no. T5168] used at 1:400 000 dilution.…”

Section: Methodsmentioning

confidence: 99%

LMNA variants cause cytoplasmic distribution of nuclear pore proteins in Drosophila and human muscle

Dialynas¹,

Flannery²,

Zirbel³

et al. 2011

Human Molecular Genetics

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Mutations in the human LMNA gene, encoding A-type lamins, give rise to laminopathies, which include several types of muscular dystrophy. Here, heterozygous sequence variants in LMNA, which result in single amino-acid substitutions, were identified in patients exhibiting muscle weakness. To assess whether the substitutions altered lamin function, we performed in vivo analyses using a Drosophila model. Stocks were generated that expressed mutant forms of the Drosophila A-type lamin modeled after each variant. Larvae were used for motility assays and histochemical staining of the body-wall muscle. In parallel, immunohistochemical analyses were performed on human muscle biopsy samples from the patients. In control flies, muscle-specific expression of the wild-type A-type lamin had no apparent affect. In contrast, expression of the mutant A-type lamins caused dominant larval muscle defects and semi-lethality at the pupal stage. Histochemical staining of larval body wall muscle revealed that the mutant A-type lamin, B-type lamins, the Sad1p, UNC-84 domain protein Klaroid and nuclear pore complex proteins were mislocalized to the cytoplasm. In addition, cytoplasmic actin filaments were disorganized, suggesting links between the nuclear lamina and the cytoskeleton were disrupted. Muscle biopsies from the patients showed dystrophic histopathology and architectural abnormalities similar to the Drosophila larvae, including cytoplasmic distribution of nuclear envelope proteins. These data provide evidence that the Drosophila model can be used to assess the function of novel LMNA mutations and support the idea that loss of cellular compartmentalization of nuclear proteins contributes to muscle disease pathogenesis.

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Section: Methodsmentioning

confidence: 99%

LMNA variants cause cytoplasmic distribution of nuclear pore proteins in Drosophila and human muscle

Dialynas¹,

Flannery²,

Zirbel³

et al. 2011

Human Molecular Genetics

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“…To determine the levels of Lamin C, protein extracts from larvae were prepared [70] and analyzed with antibodies to Lamin C (LC28.26 anti-mouse IgG) [21] used at 1∶5000 dilution. Antibodies to alpha-tubulin (anti-mouse IgG1, T5168, Sigma) were used at 1∶400,000 dilution as a control for protein loading.…”

Section: Methodsmentioning

confidence: 99%

A Comparative Study of Drosophila and Human A-Type Lamins

et al. 2009

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Nuclear intermediate filament proteins, called lamins, form a meshwork that lines the inner surface of the nuclear envelope. Lamins contain three domains: an N-terminal head, a central rod and a C-terminal tail domain possessing an Ig-fold structural motif. Lamins are classified as either A- or B-type based on structure and expression pattern. The Drosophila genome possesses two genes encoding lamins, Lamin C and lamin Dm0, which have been designated A- and B-type, respectively, based on their expression profile and structural features. In humans, mutations in the gene encoding A-type lamins are associated with a spectrum of predominantly tissue-specific diseases known as laminopathies. Linking the disease phenotypes to cellular functions of lamins has been a major challenge. Drosophila is being used as a model system to identify the roles of lamins in development. Towards this end, we performed a comparative study of Drosophila and human A-type lamins. Analysis of transgenic flies showed that human lamins localize predictably within the Drosophila nucleus. Consistent with this finding, yeast two-hybrid data demonstrated conservation of partner-protein interactions. Drosophila lacking A-type lamin show nuclear envelope defects similar to those observed with human laminopathies. Expression of mutant forms of the A-type Drosophila lamin modeled after human disease-causing amino acid substitutions revealed an essential role for the N-terminal head and the Ig-fold in larval muscle tissue. This tissue-restricted sensitivity suggests a conserved role for lamins in muscle biology. In conclusion, we show that (1) localization of A-type lamins and protein-partner interactions are conserved between Drosophila and humans, (2) loss of the Drosophila A-type lamin causes nuclear defects and (3) muscle tissue is sensitive to the expression of mutant forms of A-type lamin modeled after those causing disease in humans. These studies provide new insights on the role of lamins in nuclear biology and support Drosophila as a model for studies of human laminopathies involving muscle dysfunction.

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“…Western analysis: To determine the expression levels of LamC, proteins were extracted from third instar larvae or adults (Friedman et al 1992) and separated by size on 10-12% polyacrylamide gels, transferred to nitrocellulose membrane, and incubated with anti-LamC LC28.26 anti-mouse IgG (Riemer et al 1995) used at 1:5000-1:8000 dilution or anti-atubulin anti-mouse IgG1 [Sigma (St. Louis) no. T5168] used at 1:400,000 dilution.…”

Section: Ex296mentioning

confidence: 99%

Molecular Genetic Analysis of the Nested Drosophila melanogaster Lamin C Gene

Schulze

Curio-Penny

et al. 2005

Genetics

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Lamins are intermediate filaments that line the inner surface of the nuclear envelope, providing structural support and making contacts with chromatin. There are two types of lamins, A-and B-types, which differ in structure and expression. Drosophila possesses both lamin types, encoded by the LamC (A-type) and lamin Dm 0 (B-type) genes. LamC is nested within an intron of the essential gene ttv. We demonstrate that null mutations in LamC are lethal, and expression of a wild-type LamC transgene rescues lethality of LamC but not ttv mutants. Mutations in the human A-type lamin gene lead to diseases called laminopathies. To determine if Drosophila might serve as a useful model to study lamin biology and disease mechanisms, we generated transgenic flies expressing mutant LamC proteins modeled after human disease-causing lamins. These transgenic animals display a nuclear lamin aggregation phenotype remarkably similar to that observed when human mutant A-type lamins are expressed in mammalian cells. LamC aggregates also cause disorganization of lamin Dm 0 , indicating interdependence of both lamin types for proper lamina assembly. Taken together, these data provide the first detailed genetic analysis of the LamC gene and support using Drosophila as a model to study the role of lamins in disease.

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The urate oxidase gene of Drosophila pseudoobscura and Drosophila melanogaster: Evolutionary changes of sequence and regulation

Cited by 12 publications

References 103 publications

LMNA variants cause cytoplasmic distribution of nuclear pore proteins in Drosophila and human muscle

LMNA variants cause cytoplasmic distribution of nuclear pore proteins in Drosophila and human muscle

A Comparative Study of Drosophila and Human A-Type Lamins

Molecular Genetic Analysis of the Nested Drosophila melanogaster Lamin C Gene

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