Allele Dmel\shot³

General Information
Symbol	Dmel\shot³	Species	D. melanogaster
Name		FlyBase ID	FBal0051150
Feature type	allele	Created / Updated	2006-08-22/2006-08-22
Associated gene	Dmel\shot

Allele class	loss of function
Mutagen	diepoxybutane
Nature of the Allele
Allele class	loss of function (Lee et al., 2000, Parrish et al., 2006)
Mutagen	diepoxybutane (Kolodziej et al., 1995)
Mapped Features and Mutations
Type Symbol & Location Additional Notes References
Associated Sequence Data
DDBJ / EMBL / GenBank	DNA sequence Protein sequence Name

UniProtKB/Swiss-Prot
UniProtKB/TrEMBL

Progenitor genotype
Nature of the lesion	Statement Reference
Assay mode
Cytology
Phenotypic Data
Phenotypic Class
	lethal \| germ-line clone (Roper and Brown, 2003) reduced cell number \| somatic clone (Reuter et al., 2003) neuroanatomy defective (Lee et al., 2000, Lee and Kolodziej, 2002) neuroanatomy defective (with Df(2R)CX1) (Lee et al., 2000) neuroanatomy defective (with shot²) (Lee et al., 2000) neuroanatomy defective \| somatic clone (Reuter et al., 2003) neuroanatomy defective \| embryonic stage \| recessive (Lee et al., 2007)
Phenotype Manifest In
	embryonic dorsal trunk (Lee and Kolodziej, 2002) embryonic lateral trunk (Lee and Kolodziej, 2002) embryonic dorsal anastomosis (Lee and Kolodziej, 2002) calyx of mushroom body \| somatic clone (Reuter et al., 2003) chordotonal organ & embryo \| lateral (Lee et al., 2000) scolopidial dendrite & embryo (Lee et al., 2000) anterior fascicle (Lee et al., 2000, Lee and Kolodziej, 2002) posterior fascicle (Lee et al., 2000) oocyte (Roper and Brown, 2003) egg chamber & follicle cell \| somatic clone (Roper and Brown, 2003) mushroom body & axon (Lee and Luo, 1999) synapse & neuromuscular junction (Lohr et al., 2002) chordotonal organ & axon \| lateral (Kolodziej et al., 1995) longitudinal connective (Kolodziej et al., 1995) abdominal posterior fascicle (Lee and Kolodziej, 2002) sensory neuron (Lee and Kolodziej, 2002) ovary \| germ-line clone (Roper and Brown, 2004) oocyte \| germ-line clone (Roper and Brown, 2004) nurse cell \| ectopic \| germ-line clone (Roper and Brown, 2004) microtubule & ovary \| germ-line clone (Roper and Brown, 2004) centrosome & ovary \| germ-line clone (Roper and Brown, 2004) axon & pCC neuron (Lee et al., 2007)
Detailed Description
	Statement Reference Mutant phenotype as assayed by Ecol\lacZ^rp staining: longitudinal connectives missing. Mutant phenotype of lateral chordotonal axons includes: missing axons. (Kolodziej et al., 1995) Mushroom body neuroblast clones induced using the MARCM system exhibit drastic axon projection defects compared with wild type controls. The origin of the pathfinding errors appears to be near the dendritic projection area. In the majority of cases the abnormal axon bundles do project in a generally wild type direction, towards the midline region. Some even eventually fuse with the medial lobe, as they do in the wild type. (Lee and Luo, 1999) The lateral chordotonal sensory axons arrest at variable stages before reaching the central nervous system (CNS); 93% extend towards the intersegmental nerve (ISN) and stall or fail to reach the ISN, 7% travel a significant distance within the ISN, but none of the axons reach the CNS border. The morphology of the LCH growth cones is normal, with the growth cone adopting the general morphology of wild-type growth cones at the place where they stall. Defects in LCH dendrite morphology are seen in shot³ and shot²/shot³ mutant embryos. 90-100% of the motor axons of all three major pathways (ISN, SNa and ISNb) stall prematurely in late stage 16/stage 17 homozygous embryos. 3% of ISN axons lack the most dorsal terminal arborisation, 68% lack the 2 most dorsal terminal arborisations and 24% lack all three terminal arborisations. 1% of ISN axons stall before entering the dorsal muscle field. The SNa and ISNb motor axons stall before reaching their muscle targets. 75% of ISNb motor axons stall in the ventral muscle field (somewhere between muscle 6 and muscle 13) and 14% stall at the muscle field border. 33% of SNa axons have one branch missing (wild-type axons form two branches), 38% lack both branches and 17% stall over the ventral muscles or are absent. Some SNa and ISNb motor axon bundles fail to exit the CNS successfully or stall earlier than the muscle field entry points in shot³/Df(2R)CX1 embryos. 61% of ISNb motor axons stall in the ventral muscle field (somewhere between muscle 6 and muscle 13) and 20% stall at the muscle field border. 40% of ISN axons lack the 2 most dorsal terminal arborisations and 42% lack all three terminal arborisations. 17% of ISN axons stall before entering the dorsal muscle field. 24% of SNa axons have one branch missing (wild-type axons form two branches), 60% lack both branches and 8% stall over the ventral muscles or are absent. 6% of ISN axons lack the most dorsal terminal arborisation, 48% lack the 2 most dorsal terminal arborisations and 43% lack all three terminal arborisations in shot²/shot³ embryos. 57% of ISNb motor axons stall in the ventral muscle field (somewhere between muscle 6 and muscle 13) and 30% stall at the muscle field border. 46% of SNa axons have one branch missing (wild-type axons form two branches) and 28% lack both branches. Muscle morphology and number is normal in late stage 16 homozygous embryos. (Lee et al., 2000) Mutant embryos show defects in motor axon extension; the ISN stalls at the approximate position where it would normally make the second dorsalmost muscle contact, the SNa stalls where it would normally bifurcate and the ISNb stalls in the ventral muscle field and fails to reach muscle 12. Mutant embryos also show defects in sensory axon extension. (Lee and Kolodziej, 2002) The tracheal dorsal trunk lumen is discontinuous at 75% of anastomosis sites in stage 14 or older mutant embryos. Lateral trunk connections and all anastomoses at the dorsal midline are affected. The overall branching pattern of trachea within hemisegments appears normal. 6% of tracheal dorsal trunk connections form apparently normally and 19% are constricted relative to wild type. Fusion cells appear to be present at the dorsal trunk anastomosis sites. Microtubules are not apically located at anastomosis sites of the tracheal dorsal trunk in mutant embryos, in contrast to wild type, though they remain largely at the cell periphery. F-actin accumulates apically in the tracheal cells but the lumen does not extend through the fusion cells at the anastomosis site. (Lee and Kolodziej, 2002) Mutant mushroom body neuroblast clones examined in the adult show abnormal processes projecting out from the calyx. Mutant mushroom body neuroblast clones have a significant reduction in cell number compared to control clones. (Reuter et al., 2003) Homozygous germ-line clones produce egg chambers, but they do not progress beyond stage 7 of oogenesis. Each mutant egg chamber contains 16 nuclei, and the pattern of divisions is also normal. Fusome structure and persistence appear normal. In addition the fusome/spectrosome is still distributed asymmetrically in early stem cell/cystoblast division, with the larger portion being found in the stem cell after division. In mutant cysts all of the nuclei, including the two at the posterior of the egg chamber appear polyploid: All of the germ-line cells adopt a nurse cell fate, no oocyte is specified. The synaptonemal complex accumulates in none or two or more cells all throughout region 2a of the cyst, but this accumulation fails to refine onto one cell only and is always lost region 2b. In wild-type ovaries, microtubules are highly concentrated around the fusome from region 2a on, and in region 2b and 3 a microtubule organising centre (MTOC) is visible in the oocyte. This organisation and concentration of microtubules is lost in mutant germ-line clones; microtubules fail to concentrate around the fusome., and an MTOC does not form in region 2b or 3. the migration of the centrosomes into the oocyte normally seen in wild-type ovaries fails to occur. (Roper and Brown, 2004) shot³ mutants show strong reduction of output synapses at the NMJ, while this phenotype is not seen in the CNS. (Lohr et al., 2002) shot³ germline clones do not produce viable eggs. shot³ clones of cells are often double-layered in egg chambers from stage 6-7 onwards, with actin accumulation at the contacts between the two layers. Double-layered clones are only detected in cells overlying the oocyte, and at stage 10 of oogenesis are generally found in the posterior half of the follicle cells covering the oocyte. Microtubule levels and organisation, integrin localisation and oocyte elongation are normal in shot³ mutant cells. (Roper and Brown, 2003) In shot[3] embryos, the pCC axon aberrantly crosses the midline in 16% of CNS segments. (Lee et al., 2007)
Interactions
	Show the genetic interaction network for Enhancers & Suppressors
Phenotypic Class
Enhanced by
Statement Reference shot³ has neuroanatomy defective \| embryonic stage phenotype, enhanceable by exba[+]/exba² (Lee et al., 2007) shot³ has neuroanatomy defective \| embryonic stage phenotype, enhanceable by exba²/exba¹ (Lee et al., 2007) robo², shot³ has neuroanatomy defective \| embryonic stage phenotype, enhanceable by exba[+]/exba² (Lee et al., 2007) shot³/shot[+], sli² has neuroanatomy defective \| embryonic stage phenotype, enhanceable by exba[+]/exba² (Lee et al., 2007)
Enhancer of
Statement Reference shot³ is an enhancer of neuroanatomy defective \| embryonic stage phenotype of robo² (Lee et al., 2007) shot³/shot[+] is an enhancer of neuroanatomy defective \| embryonic stage phenotype of exba²/exba¹ (Lee et al., 2007) shot³/shot[+] is an enhancer of neuroanatomy defective \| embryonic stage phenotype of sli² (Lee et al., 2007)
Phenotype Manifest In
Enhanced by
Statement Reference shot³ has axon & pCC neuron phenotype, enhanceable by exba[+]/exba² (Lee et al., 2007) shot³ has axon & pCC neuron phenotype, enhanceable by exba²/exba¹ (Lee et al., 2007) robo², shot³ has axon & pCC neuron phenotype, enhanceable by exba[+]/exba² (Lee et al., 2007) shot³/shot[+], sli² has axon & pCC neuron phenotype, enhanceable by exba[+]/exba² (Lee et al., 2007)
Enhancer of
Statement Reference shot³ is an enhancer of axon & pCC neuron phenotype of robo² (Lee et al., 2007) shot³/shot[+] is an enhancer of axon & pCC neuron phenotype of exba²/exba¹ (Lee et al., 2007) shot³/shot[+] is an enhancer of axon & pCC neuron phenotype of sli² (Lee et al., 2007)
Additional Comments
Genetic Interactions
Statement Reference The frequency of aberrant pCC axon midline crossovers in shot[3]/+; exba[1]/exba[2] and shot[3]/shot[3]; exba[2]/+ double mutant embryos is significantly greater than in either single mutant homozygous embryos. The midline crossing phenotype is slightly more severe in shot[3]/shot[3]; exba[1]/exba[2] embryos compared to shot[3]/shot[3]; exba[2]/+ embryos, while shot[3]/+; exba[2]/+ embryos show no significant midline defects. The pCC axon midline crossing phenotype of robo[2] embryos is dramatically enhanced in robo[2]/+; exba[2]/+ embryos and in shot[3], robo[2]/+ embryos. The phenotype is further enhanced in shot[3], robo[2]/+; exba[2]/+ embryos. (Lee et al., 2007) The axon midline phenotype of sli[2]/+ embryos is enhanced in sli[2]/+; exba[2]/+ and shot[3]/+, sli[2]/+ double mutants. The phenotype is further enhanced in shot[3]/+, sli[2]/+; exba[2]/+ triple mutants. (Lee et al., 2007)
Xenogenetic Interactions
Statement Reference
Complementation & Rescue Data
Fails to complement	shot⁸ (Reuter et al., 2003) shot⁷ (Reuter et al., 2003) shot⁶ (Reuter et al., 2003)
Rescued by	shot³ is rescued by shot^{LA-ΔGAS2.Scer\UAS.T:Avic\GFP}/Scer\GAL4^btl.PS (Lee and Kolodziej, 2002) shot³ is rescued by shot^{LC.Scer\UAS.T:Avic\GFP}/Scer\GAL4^btl.PS (Lee and Kolodziej, 2002) shot³ is rescued by shot^{LA.Scer\UAS.T:Avic\GFP}/Scer\GAL4^esg-NP5130 (Lee and Kolodziej, 2002) shot³ is rescued by Scer\GAL4^btl.PS/shot^{LA.Scer\UAS.T:Avic\GFP} (Lee and Kolodziej, 2002) shot³ is rescued by shot^{LB.Scer\UAS.T:Avic\GFP}/Scer\GAL4¹⁴⁰⁷ (Lee and Kolodziej, 2002) shot³ is rescued by shot^{LA.Scer\UAS.T:Avic\GFP}/Scer\GAL4¹⁴⁰⁷ (Lee and Kolodziej, 2002, Lee et al., 2007) shot³ is rescued by shot^{LA-Δrod1.Scer\UAS.T:Avic\GFP}/Scer\GAL4¹⁴⁰⁷ (Lee and Kolodziej, 2002) shot³ is rescued by shot^{LA-ΔGAS2.Scer\UAS.T:Avic\GFP}/Scer\GAL4¹⁴⁰⁷ (Lee et al., 2007)
Partially rescued by	shot³ is partially rescued by shot^{LA-ΔEF-hand.Scer\UAS.T:Avic\GFP}/Scer\GAL4¹⁴⁰⁷ (Lee and Kolodziej, 2002, Lee et al., 2007)
Not rescued by	shot³ is not rescued by shot^{LC.Scer\UAS.T:Avic\GFP}/Scer\GAL4¹⁴⁰⁷ (Lee and Kolodziej, 2002, Lee et al., 2007) shot³ is not rescued by shot^{LC-ΔGAS2.Scer\UAS.T:Avic\GFP}/Scer\GAL4^btl.PS (Lee and Kolodziej, 2002) shot³ is not rescued by shot^{LA-ΔGAS2.Scer\UAS.T:Avic\GFP}/Scer\GAL4¹⁴⁰⁷ (Lee and Kolodziej, 2002)
Comments
Stocks ( 2 )
Bloomington	5141 w^*; P{FRT(w^hs)}G13 shot³/CyO, P{ase-lacZF:2.0}PK2
Kyoto	108072 w^*; P{FRT(w^hs)}G13 shot³/CyO, P{ase-lacZF:2.0}PK2
Notes on Origin
Discoverer

Synonyms & Secondary IDs ( 1 )
Reported As
Symbol Synonym	shot³ (Parrish et al., 2006, Christensen and Cook, 2007.5.8, Christensen et al., 2007.10.29, Lee et al., 2007, Lee et al., 2007)
Name Synonym
Secondary FlyBase IDs

References ( 14 )
Research paper	Lee et al., 2007, Development 134(9): 1767--1777 The F-actin-microtubule crosslinker Shot is a platform for Krasavietz-mediated translational regulation of midline axon repulsion. [FBrf0201378] Parrish et al., 2006, Genes Dev. 20(7): 820--835 Genome-wide analyses identify transcription factors required for proper morphogenesis of Drosophila sensory neuron dendrites. [FBrf0190556] Roper and Brown, 2004, Curr. Biol. 14(2): 99--110 A Spectraplakin Is Enriched on the Fusome and Organizes Microtubules during Oocyte Specification in Drosophila. [FBrf0167424] Roper and Brown, 2003, J. Cell Biol. 162(7): 1305--1315 Maintaining epithelial integrity: a function for gigantic spectraplakin isoforms in adherens junctions. [FBrf0167766] Lee and Kolodziej, 2002, Development 129(6): 1509--1520 The plakin Short Stop and the RhoA GTPase are required for E-cadherin-dependent apical surface remodeling during tracheal tube fusion. [FBrf0144831] Lee and Kolodziej, 2002, Development 129(5): 1195--1204 Short Stop provides an essential link between F-actin and microtubules during axon extension. [FBrf0144820] Lohr et al., 2002, J. Neurosci. 22(23): 10357--10367 Compartmentalization of central neurons in Drosophila: a new strategy of mosaic analysis reveals localization of presynaptic sites to specific segments of neurites. [FBrf0152141] Lee et al., 2000, J. Neurosci. 20(3): 1096--1108 short stop is allelic to kakapo, and encodes rod-like cytoskeletal-associated proteins required for axon extension. [FBrf0125142] Lee and Luo, 1999, Neuron 22(3): 451--461 Mosaic analysis with a repressible neurotechnique cell marker for studies of gene function in neuronal morphogenesis. [FBrf0107846] Kolodziej et al., 1995, Neuron 15(2): 273--286 Mutations that affect the length, fasciculation, or ventral orientation of specific sensory axons in the Drosophila embryo. [FBrf0083232]
Supplementary material	Lee et al., 2007, Development 134(9): Supplemental Figure 2. [FBrf0202566]
Personal communication to FlyBase	Christensen and Cook, 2007.5.8, Isolation and characterization of Df(2R)BSC307. Isolation and characterization of Df(2R)BSC307. [FBrf0199305] Christensen et al., 2007.10.29, Isolation and characterization of Df(2R)BSC383. Isolation and characterization of Df(2R)BSC383. [FBrf0200287]
Abstract	Reuter et al., 2003, Development 130(6): 1203--1213 A mosaic genetic screen for genes necessary for Drosophila mushroom body neuronal morphogenesis. [FBrf0155698]

Allele Dmel\shot3

Allele Dmel\shot³