Molecular Genetic Analysis of the Nested <i>Drosophila melanogaster</i> Lamin C Gene

Schulze, Sandra; Curio-Penny, Beatrice; Li, Yuhong; Imani, Reza; Rydberg, Lena; Geyer, Pamela; Wallrath, Lori L.

doi:10.1534/genetics.105.043208

Cited by 54 publications

(67 citation statements)

References 46 publications

(53 reference statements)

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“…Generation of transgenic stocks expressing wild type and Lamin C N was previously reported (Schulze et al, 2005;Schulze et al, 2009). The GAL4-UAS system (Brand and Perrimon, 1993; Duffy, 2002) was used to drive expression of wild type Lamin C and Lamin C N in specific tissues.…”

Section: Drosophila Stocks and Genetic Analysesmentioning

confidence: 99%

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The role ofDrosophilaLamin C in muscle function and gene expression

et al. 2010

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SUMMARYThe inner side of the nuclear envelope (NE) is lined with lamins, a meshwork of intermediate filaments that provides structural support for the nucleus and plays roles in many nuclear processes. Lamins, classified as A-or B-types on the basis of biochemical properties, have a conserved globular head, central rod and C-terminal domain that includes an Ig-fold structural motif. In humans, mutations in A-type lamins give rise to diseases that exhibit tissue-specific defects, such as Emery-Dreifuss muscular dystrophy. Drosophila is being used as a model to determine tissue-specific functions of A-type lamins in development, with implications for understanding human disease mechanisms. The GAL4-UAS system was used to express wild-type and mutant forms of Lamin C (the presumed Drosophila A-type lamin), in an otherwise wild-type background. Larval muscle-specific expression of wild type Drosophila Lamin C caused no overt phenotype. By contrast, larval muscle-specific expression of a truncated form of Lamin C lacking the N-terminal head (Lamin C N) caused muscle defects and semi-lethality, with adult 'escapers' possessing malformed legs. The leg defects were due to a lack of larval muscle function and alterations in hormoneregulated gene expression. The consequences of Lamin C association at a gene were tested directly by targeting a Lamin C DNAbinding domain fusion protein upstream of a reporter gene. Association of Lamin C correlated with localization of the reporter gene at the nuclear periphery and gene repression. These data demonstrate connections among the Drosophila A-type lamin, hormone-induced gene expression and muscle function. DEVELOPMENT 3068 lethality when expressed in larval muscle but not in other tissues (Schulze et al., 2009). By contrast, alterations in the rod domain cause abnormal lamin aggregation within the nucleus but not lethality. Collectively, our studies support a role for the N-terminal head and C-terminal globular domain in larval muscle function.To better understand the role of the N-terminal head domain, we generated transgenic flies expressing a mutant form of Lamin C lacking the first 42 amino acids (Lamin C N) (Schulze et al., 2009). This mutant form localizes to the NE and causes semilethality when expressed in muscle but not in non-muscle tissues (Schulze et al., 2009). Here, we demonstrate that lethality is associated with larval body wall muscle defects that include nuclear and cytoplasmic abnormalities. 'Adult escapers' expressing Lamin C N have leg defects that are consistent with a loss of muscle function and disruptions in ecdysone hormone signaling.Collectively, these studies demonstrate a role for the N-terminal head domain of Lamin C in muscle function and gene expression. MATERIALS AND METHODS Drosophila stocks and genetic analysesDrosophila stocks were raised at room temperature on standard sucrose and cornmeal medium (Shaffer et al., 1994). Generation of transgenic stocks expressing wild type and Lamin C N was previously reported (Schulze et al., 2005;Schulze ...

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Section: Drosophila Stocks and Genetic Analysesmentioning

confidence: 99%

“…Drosophila is the only invertebrate model organism known to possess the two types of lamins found in humans (Klapper et al, 1997;Riemer et al, 1995;Schulze et al, 2005). The Drosophila genome possesses two lamin genes: Lamin C, proposed to encode the A-type lamin, and Lamin Dm 0 , the B-type lamin (Riemer et al, 1995).…”

Section: Introductionmentioning

confidence: 99%

The role ofDrosophilaLamin C in muscle function and gene expression

et al. 2010

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“…Live imaging of embryos expressing mRFP-Nup107 and GFP-Lamin C (from a protein trap transgenic line described in Schulze et al, 2005) ( Figure 3A and Supplemental Movie 2) revealed that the GFP-Lamin C signal at the NE declined more progressively in prometaphase than mRFP-Nup107 (see quantifications in Figure 4A). GFP-Lamin C persisted at the SE into metaphase, where it surrounded the diffuse mRFP-Nup107 signal.…”

Section: Ne and Npc Dynamics During Drosophila Embryonic Mitosismentioning

confidence: 99%

In Vivo Dynamics ofDrosophilaNuclear Envelope Components

Katsani

Karess

Dostatni

et al. 2008

MBoC

118

128

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Nuclear pore complexes (NPCs) are multisubunit protein entities embedded into the nuclear envelope (NE). Here, we examine the in vivo dynamics of the essential Drosophila nucleoporin Nup107 and several other NE-associated proteins during NE and NPCs disassembly and reassembly that take place within each mitosis. During both the rapid mitosis of syncytial embryos and the more conventional mitosis of larval neuroblasts, Nup107 is gradually released from the NE, but it remains partially confined to the nuclear (spindle) region up to late prometaphase, in contrast to nucleoporins detected by wheat germ agglutinin and lamins. We provide evidence that in all Drosophila cells, a structure derived from the NE persists throughout metaphase and early anaphase. Finally, we examined the dynamics of the spindle checkpoint proteins Mad2 and Mad1. During mitotic exit, Mad2 and Mad1 are actively imported back from the cytoplasm into the nucleus after the NE and NPCs have reformed, but they reassociate with the NE only later in G1, concomitantly with the recruitment of the basket nucleoporin Mtor (the Drosophila orthologue of vertebrate Tpr). Surprisingly, Drosophila Nup107 shows no evidence of localization to kinetochores, despite the demonstrated importance of this association in mammalian cells. INTRODUCTIONIn eukaryotes, the nuclear envelope (NE) defines the limits between the nucleus and the cytoplasm. The outer NE membrane is considered to be structurally and functionally part of the endoplasmic reticulum network, whereas the inner membrane, with its distinct protein composition, provides anchoring points for the chromatin and nuclear lamina. Nuclear pore complexes (NPCs) are embedded at the points of fusion between the inner and outer NE membrane and represent the sole channels of transport across the NE. NPCs are composed of multiple copies of ϳ30 different proteins termed nucleoporins (Nups), most of which are organized into subcomplexes that associate with each other to build up the mature NPCs (for reviews, see Hetzer et al., 2005;Schwartz, 2005;Lim and Fahrenkrog, 2006;Tran and Wente, 2006). During cell division, the NE and NPCs are subjected to major rearrangements. However, the extent to which the NE and NPCs disassemble at mitotic entry varies among organisms (for reviews, see Margalit et al., 2005;Prunuske and Ullman, 2006). Unlike in most yeast and fungi, characterized by a "closed mitosis," NE disassembly is required in animal cells to allow spindle microtubule access to chromosomes. In vertebrates, cell division leads to complete NE breakdown at the prophase-prometaphase transition. During this "open mitosis," integral membrane proteins of the NE and the soluble subcomplexes of the NPCs redistribute throughout the endoplasmic reticulum and the mitotic cytoplasm (for reviews, see Hetzer et al., 2005;Margalit et al., 2005;Prunuske and Ullman, 2006). In Drosophila and Caenorhabditis elegans embryos, however, the NE only partially disassembles near spindle poles in early mitosis. NPCs disassemble during prometapha...

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“…Immunolocalization studies demonstrate that dMAN1, Bocksbeutel, and Otefin are enriched at the nuclear envelope (Padan et al 1990;Ashery-Padan et al 1997a,b;Wagner et al 2004aWagner et al , 2006Jiang et al 2008). Drosophila provides an excellent model for studies of the developmental roles of the nuclear lamina proteins, as homologs of many vertebrate lamina proteins have been identified, including A-and B-type lamins (lamin C and lamin Dm 0 , respectively) (Fisher et al 1982;Lenz-Bohme et al 1997;Osouda et al 2005;Schulze et al 2005), Drosophila (d)BAF (Furukawa et al 2003), the lamin B receptor (Wagner et al 2004b), and proteins in the linker of nucleoskeleton and cytoskeleton (LINC) complex (Starr and Fischer 2005;Stewart et al 2007).…”

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confidence: 99%

Tissue-Specific Defects Are Caused by Loss of the Drosophila MAN1 LEM Domain Protein

Pinto¹,

Wilmington

Hornick

et al. 2008

Genetics

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The nuclear lamina represents a protein network required for nuclear structure and function. One family of lamina proteins is defined by an $40-aa LAP2, Emerin, and MAN1 (LEM) domain (LEM-D) that binds the nonspecific DNA-binding protein, barrier-to-autointegration factor (BAF). Through interactions with BAF, LEM-D proteins serve as a bridge between chromosomes and the nuclear envelope. Mutations in genes encoding LEM-D proteins cause human laminopathies that are associated with tissue-restricted pathologies. Drosophila has five genes that encode proteins with LEM homology. Using yeast two-hybrid analyses, we demonstrate that four encode proteins that bind Drosophila (d)BAF. In addition to dBAF, dMAN1 associates with lamins, the LEM-D protein Bocksbeutel, and the receptor-regulated Smads, demonstrating parallel protein interactions with vertebrate homologs. P-element mobilization was used to generate null dMAN1 alleles. These mutants showed decreased viability, with surviving adults displaying male sterility, decreased female fertility, wing patterning and positioning defects, flightlessness, and locomotion difficulties that became more severe with age. Increased phospho-Smad staining in dMAN1 mutant wing discs is consistent with a role in transforming growth factor (TGF)-b/bone morphogenic protein (BMP) signaling. The tissuespecific, age-enhanced dMAN1 mutant phenotypes are reminiscent of human laminopathies, suggesting that studies in Drosophila will provide insights into lamina dysfunction associated with disease.

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Molecular Genetic Analysis of the Nested Drosophila melanogaster Lamin C Gene

Cited by 54 publications

References 46 publications

The role ofDrosophilaLamin C in muscle function and gene expression

The role ofDrosophilaLamin C in muscle function and gene expression

In Vivo Dynamics ofDrosophilaNuclear Envelope Components

Tissue-Specific Defects Are Caused by Loss of the Drosophila MAN1 LEM Domain Protein

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