Characterization of a new oda3 allele, oda3-6, defective in assembly of the outer dynein arm-docking complex in Chlamydomonas reinhardtii

Abstract

We have used an insertional mutagenesis approach to generate new C. reinhardtii motility mutants. Of 56 mutants isolated, one is a new allele at the ODA3 locus, called oda3-6. Similar to the previously characterized oda3 alleles, oda3-6 has a slow-jerky swimming phenotype and reduced swimming speed. The oda3-6 mutant fails to assemble the outer dynein arm motor and outer dynein arm—docking complex (ODA-DC) in the ciliary axoneme due to an insertion in the 5’ end of the DCC1 gene, which encodes the DC1 subunit of the ODA-DC. Transformation of oda3-6 with the wild-type DCC1 gene rescues the mutant swimming phenotype and restores assembly of the ODA-DC and the outer dynein arm in the cilium. This is the first oda3 mutant to be characterized at the molecular level and is likely to be very useful for further analysis of DC1 function.

Structure of the Decorated Ciliary Doublet Microtubule

“…The authors declare no competing interests. Blanchon et al, (2012), Bragina et al, (2019), Brown et al, (2017), Davis et al, (2019), Gegg et al, (2014), Geremek et al, (2008), Ghandourah and Dell, (2018), Hjeij et al, (2014), Knowles et al, (2013), Li et al, (2019a), Lorenzetti et al, (2004), Meng et al, (2014), Merveille et al, (2011), Mira et al, (2004), Mizuno et al, (2016), Narasimhan et al, (2015), Onoufriadis et al, (2013), Perles et al, (2012), Reish et al, (2016), Sigg et al, (2017), Subaran et al, (2015), Sung et al, (2016), Ta-Shma et al, (2015, Vadnais et al, (2014), Wilson et al, (2010a), Wilson et al, (2010b), and Zhang et al, (2019). Sung, Y.H., Baek, I.-J., Kim, Y.H., Gho, Y.S., Oh, S.P., Lee, Y.J., and Lee, H.-W. (2016).…”

PKA, PP1, and DC1 phosphorylation mediate alcohol-induced ciliary dysfunction in Chlamydomonas reinhardtii

“…A 1.8-kb EcoRV-PvuII restriction fragment from the pHyg3 plasmid containing the hygromycin-resistance cassette (Berthold, Schmitt, & Mages, 2002) was cloned into the EcoRV site of the DCC1 pCS3.3 plasmid (Brown et al, 2017) to make plasmid pMM4.39.…”

The IDA3 adapter, required for intraflagellar transport of I1 dynein, is regulated by ciliary length

“…Defective motility can arise from the failure to assemble dynein arms ( Kamiya and Yagi, 2014 ). Proper assembly of axonemal dynein arms requires not only the structural components, such as dynein heavy, intermediate, and light chains, but also factors extrinsic to the dynein for cytoplasmic assembly ( Iomini et al , 2009 ; Kobayashi and Takeda, 2012 ), intraflagellar transport (IFT) ( Hou et al , 2007 ; Ahmed et al , 2008 ; Viswanadha et al , 2014 ; Desai et al , 2015 ; Lechtreck, 2015 ; Hou and Witman, 2017 ; Taschner et al , 2017 ), and docking in the axoneme ( Owa et al , 2014 ; Brown et al , 2017 ). We are only beginning to understand the key proteins and mechanisms required to assemble and transport ciliary dyneins, as well as other complexes in the axoneme ( Lechtreck et al , 2017 ).…”