Journal
CELL REPORTS
Volume 27, Issue 8, Pages 2459-+Publisher
CELL PRESS
DOI: 10.1016/j.celrep.2019.04.087
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Funding
- NSF GRFP grant [1518767]
- [P20 GM104936-09]
- [1R35GM128702-01]
- Direct For Education and Human Resources
- Division Of Graduate Education [1518767] Funding Source: National Science Foundation
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The unicellular green alga Chlamydomonas reinhardtii is a biflagellated cell with two actin genes: one encoding a conventional actin (IDA5) and the other encoding a divergent novel actin-like protein (NAP1). Here, we probe how actin redundancy contributes to flagellar assembly. Disrupting a single actin allows complete flagellar assembly. However, when disrupting both actins using latrunculin B (LatB) treatment on the napl mutant background, we find that actins are necessary for flagellar growth from newly synthesized limiting flagellar proteins. Under total actin disruption, transmission electron microscopy identified an accumulation of Golgiadjacent vesicles. We also find that there is a mislocalization of a key transition zone gating and ciliopathy protein, NPHP-4. Our experiments demonstrate that each stage of flagellar biogenesis requires redundant actin function to varying degrees, with an absolute requirement for these actins in transport of Golgi-adjacent vesicles and flagellar incorporation of newly synthesized proteins.
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