Identification of Proteins Required for Precise Positioning of Apc2 in Dendrites
In Drosophila neurons, uniform minus-end-out polarity in dendrites is maintained in part by kinesin-2-mediated steering of growing microtubules at branch points. Apc links the kinesin motor to growing microtubule plus ends and Apc2 recruits Apc to branch points where it functions. Because Apc2 acts...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Oxford University Press
2018-05-01
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| Series: | G3: Genes, Genomes, Genetics |
| Subjects: | |
| Online Access: | http://g3journal.org/lookup/doi/10.1534/g3.118.200205 |
| _version_ | 1818894154820747264 |
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| author | Alexis T. Weiner Dylan Y. Seebold Nick L. Michael Michelle Guignet Chengye Feng Brandon Follick Brandon A. Yusko Nathan P. Wasilko Pedro Torres-Gutierrez Melissa M. Rolls |
| author_facet | Alexis T. Weiner Dylan Y. Seebold Nick L. Michael Michelle Guignet Chengye Feng Brandon Follick Brandon A. Yusko Nathan P. Wasilko Pedro Torres-Gutierrez Melissa M. Rolls |
| author_sort | Alexis T. Weiner |
| collection | DOAJ |
| description | In Drosophila neurons, uniform minus-end-out polarity in dendrites is maintained in part by kinesin-2-mediated steering of growing microtubules at branch points. Apc links the kinesin motor to growing microtubule plus ends and Apc2 recruits Apc to branch points where it functions. Because Apc2 acts to concentrate other steering proteins to branch points, we wished to understand how Apc2 is targeted. From an initial broad candidate RNAi screen, we found Miro (a mitochondrial transport protein), Ank2, Axin, spastin and Rac1 were required to position Apc2-GFP at dendrite branch points. YFP-Ank2-L8, Axin-GFP and mitochondria also localized to branch points suggesting the screen identified relevant proteins. By performing secondary screens, we found that energy production by mitochondria was key for Apc2-GFP positioning and spastin acted upstream of mitochondria. Ank2 seems to act independently from other players, except its membrane partner, Neuroglian (Nrg). Rac1 likely acts through Arp2/3 to generate branched actin to help recruit Apc2-GFP. Axin can function in a variety of wnt signaling pathways, one of which includes heterotrimeric G proteins and Frizzleds. Knockdown of Gαs, Gαo, Fz and Fz2, reduced targeting of Apc2 and Axin to branch points. Overall our data suggest that mitochondrial energy production, Nrg/Ank2, branched actin generated by Arp2/3 and Fz/G proteins/Axin function as four modules that control localization of the microtubule regulator Apc2 to its site of action in dendrite branch points. |
| first_indexed | 2024-12-19T18:23:59Z |
| format | Article |
| id | doaj.art-4e7685699f244454acd1ed5fa12fdb22 |
| institution | Directory Open Access Journal |
| issn | 2160-1836 |
| language | English |
| last_indexed | 2024-12-19T18:23:59Z |
| publishDate | 2018-05-01 |
| publisher | Oxford University Press |
| record_format | Article |
| series | G3: Genes, Genomes, Genetics |
| spelling | doaj.art-4e7685699f244454acd1ed5fa12fdb222022-12-21T20:10:53ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362018-05-01851841185310.1534/g3.118.20020542Identification of Proteins Required for Precise Positioning of Apc2 in DendritesAlexis T. WeinerDylan Y. SeeboldNick L. MichaelMichelle GuignetChengye FengBrandon FollickBrandon A. YuskoNathan P. WasilkoPedro Torres-GutierrezMelissa M. RollsIn Drosophila neurons, uniform minus-end-out polarity in dendrites is maintained in part by kinesin-2-mediated steering of growing microtubules at branch points. Apc links the kinesin motor to growing microtubule plus ends and Apc2 recruits Apc to branch points where it functions. Because Apc2 acts to concentrate other steering proteins to branch points, we wished to understand how Apc2 is targeted. From an initial broad candidate RNAi screen, we found Miro (a mitochondrial transport protein), Ank2, Axin, spastin and Rac1 were required to position Apc2-GFP at dendrite branch points. YFP-Ank2-L8, Axin-GFP and mitochondria also localized to branch points suggesting the screen identified relevant proteins. By performing secondary screens, we found that energy production by mitochondria was key for Apc2-GFP positioning and spastin acted upstream of mitochondria. Ank2 seems to act independently from other players, except its membrane partner, Neuroglian (Nrg). Rac1 likely acts through Arp2/3 to generate branched actin to help recruit Apc2-GFP. Axin can function in a variety of wnt signaling pathways, one of which includes heterotrimeric G proteins and Frizzleds. Knockdown of Gαs, Gαo, Fz and Fz2, reduced targeting of Apc2 and Axin to branch points. Overall our data suggest that mitochondrial energy production, Nrg/Ank2, branched actin generated by Arp2/3 and Fz/G proteins/Axin function as four modules that control localization of the microtubule regulator Apc2 to its site of action in dendrite branch points.http://g3journal.org/lookup/doi/10.1534/g3.118.200205dendriteDrosophilaprotein localization |
| spellingShingle | Alexis T. Weiner Dylan Y. Seebold Nick L. Michael Michelle Guignet Chengye Feng Brandon Follick Brandon A. Yusko Nathan P. Wasilko Pedro Torres-Gutierrez Melissa M. Rolls Identification of Proteins Required for Precise Positioning of Apc2 in Dendrites G3: Genes, Genomes, Genetics dendrite Drosophila protein localization |
| title | Identification of Proteins Required for Precise Positioning of Apc2 in Dendrites |
| title_full | Identification of Proteins Required for Precise Positioning of Apc2 in Dendrites |
| title_fullStr | Identification of Proteins Required for Precise Positioning of Apc2 in Dendrites |
| title_full_unstemmed | Identification of Proteins Required for Precise Positioning of Apc2 in Dendrites |
| title_short | Identification of Proteins Required for Precise Positioning of Apc2 in Dendrites |
| title_sort | identification of proteins required for precise positioning of apc2 in dendrites |
| topic | dendrite Drosophila protein localization |
| url | http://g3journal.org/lookup/doi/10.1534/g3.118.200205 |
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