Abstract
Our previous genetic analysis of synaptic mechanisms in Drosophila identified a temperature-sensitive paralytic mutant of the voltage-gated calcium channel alpha1 subunit gene, cacophony (cac). Electrophysiological studies in this mutant, designated cac(TS2), indicated cac encodes a primary calcium channel alpha1 subunit functioning in neurotransmitter release. To further examine the functions and interactions of cac-encoded calcium channels, a genetic screen was performed to isolate new mutations that modify the cac(TS2) paralytic phenotype. The screen recovered 10 mutations that enhance or suppress cac(TS2), including second-site mutations in cac (intragenic modifiers) as well as mutations mapping to other genes (extragenic modifiers). Here we report molecular characterization of three intragenic modifiers and examine the consequences of these mutations for temperature-sensitive behavior, synaptic function, and processing of cac pre-mRNAs. These mutations may further define the structural basis of calcium channel alpha1 subunit function in neurotransmitter release.
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- Chan Betty, Villella Adriana, Funes Pablo, Hall Jeffrey C. Courtship and other behaviors affected by a heat-sensitive, molecularly novel mutation in the cacophony calcium-channel gene of Drosophila. Genetics. 2002 Sep;162(1):135–153. doi: 10.1093/genetics/162.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
- DeMaria C. D., Soong T. W., Alseikhan B. A., Alvania R. S., Yue D. T. Calmodulin bifurcates the local Ca2+ signal that modulates P/Q-type Ca2+ channels. Nature. 2001 May 24;411(6836):484–489. doi: 10.1038/35078091. [DOI] [PubMed] [Google Scholar]
- Dellinger B., Felling R., Ordway R. W. Genetic modifiers of the Drosophila NSF mutant, comatose, include a temperature-sensitive paralytic allele of the calcium channel alpha1-subunit gene, cacophony. Genetics. 2000 May;155(1):203–211. doi: 10.1093/genetics/155.1.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dove L. S., Abbott L. C., Griffith W. H. Whole-cell and single-channel analysis of P-type calcium currents in cerebellar Purkinje cells of leaner mutant mice. J Neurosci. 1998 Oct 1;18(19):7687–7699. doi: 10.1523/JNEUROSCI.18-19-07687.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gautam M., Tanouye M. A. Alteration of potassium channel gating: molecular analysis of the Drosophila Sh5 mutation. Neuron. 1990 Jul;5(1):67–73. doi: 10.1016/0896-6273(90)90034-d. [DOI] [PubMed] [Google Scholar]
- Grigliatti T. A., Hall L., Rosenbluth R., Suzuki D. T. Temperature-sensitive mutations in Drosophila melanogaster. XIV. A selection of immobile adults. Mol Gen Genet. 1973 Jan 24;120(2):107–114. doi: 10.1007/BF00267238. [DOI] [PubMed] [Google Scholar]
- Horn R. A new twist in the saga of charge movement in voltage-dependent ion channels. Neuron. 2000 Mar;25(3):511–514. doi: 10.1016/s0896-6273(00)81055-7. [DOI] [PubMed] [Google Scholar]
- Ikeda S. R., Dunlap K. Voltage-dependent modulation of N-type calcium channels: role of G protein subunits. Adv Second Messenger Phosphoprotein Res. 1999;33:131–151. doi: 10.1016/s1040-7952(99)80008-1. [DOI] [PubMed] [Google Scholar]
- Ino M., Yoshinaga T., Wakamori M., Miyamoto N., Takahashi E., Sonoda J., Kagaya T., Oki T., Nagasu T., Nishizawa Y. Functional disorders of the sympathetic nervous system in mice lacking the alpha 1B subunit (Cav 2.2) of N-type calcium channels. Proc Natl Acad Sci U S A. 2001 Apr 10;98(9):5323–5328. doi: 10.1073/pnas.081089398. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jun K., Piedras-Rentería E. S., Smith S. M., Wheeler D. B., Lee S. B., Lee T. G., Chin H., Adams M. E., Scheller R. H., Tsien R. W. Ablation of P/Q-type Ca(2+) channel currents, altered synaptic transmission, and progressive ataxia in mice lacking the alpha(1A)-subunit. Proc Natl Acad Sci U S A. 1999 Dec 21;96(26):15245–15250. doi: 10.1073/pnas.96.26.15245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kanevsky M., Aldrich R. W. Determinants of voltage-dependent gating and open-state stability in the S5 segment of Shaker potassium channels. J Gen Physiol. 1999 Aug;114(2):215–242. doi: 10.1085/jgp.114.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawasaki F., Felling R., Ordway R. W. A temperature-sensitive paralytic mutant defines a primary synaptic calcium channel in Drosophila. J Neurosci. 2000 Jul 1;20(13):4885–4889. doi: 10.1523/JNEUROSCI.20-13-04885.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawasaki F., Mattiuz A. M., Ordway R. W. Synaptic physiology and ultrastructure in comatose mutants define an in vivo role for NSF in neurotransmitter release. J Neurosci. 1998 Dec 15;18(24):10241–10249. doi: 10.1523/JNEUROSCI.18-24-10241.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawasaki F., Ordway R. W. The Drosophila NSF protein, dNSF1, plays a similar role at neuromuscular and some central synapses. J Neurophysiol. 1999 Jul;82(1):123–130. doi: 10.1152/jn.1999.82.1.123. [DOI] [PubMed] [Google Scholar]
- Kawasaki Fumiko, Collins Stephen C., Ordway Richard W. Synaptic calcium-channel function in Drosophila: analysis and transformation rescue of temperature-sensitive paralytic and lethal mutations of cacophony. J Neurosci. 2002 Jul 15;22(14):5856–5864. doi: 10.1523/JNEUROSCI.22-14-05856.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kraus R. L., Sinnegger M. J., Glossmann H., Hering S., Striessnig J. Familial hemiplegic migraine mutations change alpha1A Ca2+ channel kinetics. J Biol Chem. 1998 Mar 6;273(10):5586–5590. doi: 10.1074/jbc.273.10.5586. [DOI] [PubMed] [Google Scholar]
- Kühn F. J., Greeff N. G. Movement of voltage sensor S4 in domain 4 is tightly coupled to sodium channel fast inactivation and gating charge immobilization. J Gen Physiol. 1999 Aug;114(2):167–183. doi: 10.1085/jgp.114.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lorenzon N. M., Beam K. G. Calcium channelopathies. Kidney Int. 2000 Mar;57(3):794–802. doi: 10.1046/j.1523-1755.2000.00917.x. [DOI] [PubMed] [Google Scholar]
- Lorenzon N. M., Lutz C. M., Frankel W. N., Beam K. G. Altered calcium channel currents in Purkinje cells of the neurological mutant mouse leaner. J Neurosci. 1998 Jun 15;18(12):4482–4489. doi: 10.1523/JNEUROSCI.18-12-04482.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsushita Kaori, Wakamori Minoru, Rhyu Im Joo, Arii Tatsuo, Oda Sen-Ichi, Mori Yasuo, Imoto Keiji. Bidirectional alterations in cerebellar synaptic transmission of tottering and rolling Ca2+ channel mutant mice. J Neurosci. 2002 Jun 1;22(11):4388–4398. doi: 10.1523/JNEUROSCI.22-11-04388.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mori Y., Wakamori M., Oda S., Fletcher C. F., Sekiguchi N., Mori E., Copeland N. G., Jenkins N. A., Matsushita K., Matsuyama Z. Reduced voltage sensitivity of activation of P/Q-type Ca2+ channels is associated with the ataxic mouse mutation rolling Nagoya (tg(rol)). J Neurosci. 2000 Aug 1;20(15):5654–5662. doi: 10.1523/JNEUROSCI.20-15-05654.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ophoff R. A., Terwindt G. M., Vergouwe M. N., van Eijk R., Oefner P. J., Hoffman S. M., Lamerdin J. E., Mohrenweiser H. W., Bulman D. E., Ferrari M. Familial hemiplegic migraine and episodic ataxia type-2 are caused by mutations in the Ca2+ channel gene CACNL1A4. Cell. 1996 Nov 1;87(3):543–552. doi: 10.1016/s0092-8674(00)81373-2. [DOI] [PubMed] [Google Scholar]
- Peixoto A. A., Smith L. A., Hall J. C. Genomic organization and evolution of alternative exons in a Drosophila calcium channel gene. Genetics. 1997 Apr;145(4):1003–1013. doi: 10.1093/genetics/145.4.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pietrobon Daniela. Calcium channels and channelopathies of the central nervous system. Mol Neurobiol. 2002 Feb;25(1):31–50. doi: 10.1385/MN:25:1:031. [DOI] [PubMed] [Google Scholar]
- Ramaswami M., Rao S., van der Bliek A., Kelly R. B., Krishnan K. S. Genetic studies on dynamin function in Drosophila. J Neurogenet. 1993 Dec;9(2):73–87. doi: 10.3109/01677069309083451. [DOI] [PubMed] [Google Scholar]
- Reenan R. A. The RNA world meets behavior: A-->I pre-mRNA editing in animals. Trends Genet. 2001 Feb;17(2):53–56. doi: 10.1016/s0168-9525(00)02169-7. [DOI] [PubMed] [Google Scholar]
- Saegusa H., Kurihara T., Zong S., Minowa O., Kazuno A., Han W., Matsuda Y., Yamanaka H., Osanai M., Noda T. Altered pain responses in mice lacking alpha 1E subunit of the voltage-dependent Ca2+ channel. Proc Natl Acad Sci U S A. 2000 May 23;97(11):6132–6137. doi: 10.1073/pnas.100124197. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salkoff L., Kelly L. Temperature-induced seizure and frequency-dependent neuromuscular block in a ts mutant of Drosophila. Nature. 1978 May 11;273(5658):156–158. doi: 10.1038/273156a0. [DOI] [PubMed] [Google Scholar]
- Siddiqi O., Benzer S. Neurophysiological defects in temperature-sensitive paralytic mutants of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3253–3257. doi: 10.1073/pnas.73.9.3253. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith L. A., Peixoto A. A., Hall J. C. RNA editing in the Drosophila DMCA1A calcium-channel alpha 1 subunit transcript. J Neurogenet. 1998 Nov;12(4):227–240. doi: 10.3109/01677069809108560. [DOI] [PubMed] [Google Scholar]
- Smith L. A., Peixoto A. A., Kramer E. M., Villella A., Hall J. C. Courtship and visual defects of cacophony mutants reveal functional complexity of a calcium-channel alpha1 subunit in Drosophila. Genetics. 1998 Jul;149(3):1407–1426. doi: 10.1093/genetics/149.3.1407. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith L. A., Wang X., Peixoto A. A., Neumann E. K., Hall L. M., Hall J. C. A Drosophila calcium channel alpha1 subunit gene maps to a genetic locus associated with behavioral and visual defects. J Neurosci. 1996 Dec 15;16(24):7868–7879. doi: 10.1523/JNEUROSCI.16-24-07868.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Suzuki D. T., Grigliatti T., Williamson R. Temperature-sensitive mutations in Drosophila melanogaster. VII. A mutation (para-ts) causing reversible adult paralysis. Proc Natl Acad Sci U S A. 1971 May;68(5):890–893. doi: 10.1073/pnas.68.5.890. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Varadi G., Strobeck M., Koch S., Caglioti L., Zucchi C., Palyi G. Molecular elements of ion permeation and selectivity within calcium channels. Crit Rev Biochem Mol Biol. 1999;34(3):181–214. doi: 10.1080/10409239991209264. [DOI] [PubMed] [Google Scholar]
- Von Schilcher F. The behavior of cacophony, a courtship song mutant in Drosophila melanogaster. Behav Biol. 1976 Jun;17(2):187–196. doi: 10.1016/s0091-6773(76)90444-2. [DOI] [PubMed] [Google Scholar]
- Wakamori M., Yamazaki K., Matsunodaira H., Teramoto T., Tanaka I., Niidome T., Sawada K., Nishizawa Y., Sekiguchi N., Mori E. Single tottering mutations responsible for the neuropathic phenotype of the P-type calcium channel. J Biol Chem. 1998 Dec 25;273(52):34857–34867. doi: 10.1074/jbc.273.52.34857. [DOI] [PubMed] [Google Scholar]
- Zhang Yi, Mori Mayra, Burgess Daniel L., Noebels Jeffrey L. Mutations in high-voltage-activated calcium channel genes stimulate low-voltage-activated currents in mouse thalamic relay neurons. J Neurosci. 2002 Aug 1;22(15):6362–6371. doi: 10.1523/JNEUROSCI.22-15-06362.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]