Generation and characterization of new alleles of quiver (qvr) that encodes an extracellular modulator of the Shaker potassium channel

Abstract

Our earlier genetic screen uncovered a paraquat-sensitive leg-shaking mutant quiver¹ (qvr¹), whose gene product interacts with the Shaker (Sh) K⁺ channel. We also mapped the qvr locus to EY04063 and noticed altered day–night activity patterns in these mutants. Such circadian behavioral defects were independently reported by another group, who employed the qvr¹ allele we supplied them, and attributed the extreme restless phenotype of EY04063 to the qvr gene. However, their report adopted a new noncanonical gene name sleepless (sss) for qvr. In addition to qvr¹ and qvr^EY, our continuous effort since the early 2000s generated a number of novel recessive qvr alleles, including ethyl methanesulfonate (EMS)-induced mutations qvr² and qvr³, and P-element excision lines qvr^ip6 (imprecise jumpout), qvr^rv7, and qvr^rv9 (revertants) derived from qvr^EY. Distinct from the original intron-located qvr¹ allele that generates abnormal-sized mRNAs, qvr², and qvr³ had their lesion sites in exons 6 and 7, respectively, producing nearly normal-sized mRNA products. A set of RNA-editing sites are nearby the lesion sites of qvr³ and qvr^EY on exon 7. Except for the revertants, all qvr alleles display a clear ether-induced leg-shaking phenotype just like Sh, and weakened climbing abilities to varying degrees. Unlike Sh, all shaking qvr alleles (except for qvr^f01257) displayed a unique activity-dependent enhancement in excitatory junction potentials (EJPs) at larval neuromuscular junctions (NMJs) at very low stimulus frequencies, with qvr^EY displaying the largest EJP and more significant NMJ overgrowth than other alleles. Our detailed characterization of a collection of qvr alleles helps to establish links between novel molecular lesions and different behavioral and physiological consequences, revealing how modifications of the qvr gene lead to a wide spectrum of phenotypes, including neuromuscular hyperexcitability, defective motor ability and activity-rest cycles.

Keywords:

• qvr/sss
• Shaker
• sleepless
• RNA editing
• Ly-6/neurotoxin superfamily
• synaptic plasticity
• potassium current inactivation
• ether-induced leg shaking

Acknowledgements

We thank Mr. Anthony McGregor, Mr. Timothy Patience, and Dr. Yu Li for assistance in manuscript preparation and proofreading.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was partially supported by NIH grants NS26528, GM80255, AG047612, and AG051513 (USA), as well as a Natural Sciences and Engineering Research Council Discovery Grant (Canada).