Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| gene (Drosophila melanogaster) |
BicD | doi: 10.7554/ eLife.10140 (Del Castillo et al., 2015) |
CG6605 | |
| gene (Drosophila melanogaster) |
Dlic | Thomas Hays lab (University of Minnesota); doi:10.1091/mbc. E08-05-0483 (Mische et al., 2008) |
CG1938 | |
| gene (Rattus norvegicus) |
Inositol trisphosphate 3-kinase A (lTPKA) |
doi:10.1091/mbc. E09-01-0083 (Johnson and Schell, 2009); doi:10.1016 /j. ydbio.2014.06.022 (Spracklen et al., 2014) |
Gene ID: 81677 | |
| gene (Pyrococcus furiosus) |
PfV | Addgene plasmid # 116933; doi:10.1016 /j.cell. 2018.05.042 (Delarue et al., 2018) |
GenBank: AB214633.1; RRID:Addgene_116933 |
|
| gene (Physcomitrella patens) |
Kin14VIb | Gohta Goshima lab; doi:10.1038/NPLANTS.2015.87 (Jonsson et al., 2015) |
||
| genetic reagent (Drosophila melanogaster) |
mat αtub-Gal4[V37] | Bloomington Drosophila Stock Center |
BDSC: 7063; FlyBase ID: FBti0016914 |
|
| genetic reagent (Drosophila melanogaster) |
Act5C-Gal4 | Bloomington Drosophila Stock Center |
BDSC: 3954; FlyBase ID: FBti0012292 |
|
| genetic reagent (Drosophila melanogaster) |
nos-Gal4-VP16 | Edwin Ferguson lab (University of Chicago); doi: 10.1016 / s0960-9822(98)70091-0 (Van Doren et al., 1998) |
||
| genetic reagent (Drosophila melanogaster) |
UAS-Dhc64C-RNAi | Bloomington Drosophila Stock Center |
BDSC: 36583; FlyBase ID: FBti0146459; TRiP.GL00543 |
|
| genetic reagent (Drosophila melanogaster) |
UAS-Dhc64C-RNAi | Bloomington Drosophila Stock Center |
BDSC: 36698; FlyBase ID: FBti0146710; TRiP.HMS01587 |
|
| genetic reagent (Drosophila melanogaster) |
UASp-Gl∆C | Thomas Hays lab (University of Minnesota); doi:10.1091/mbc. E06-10-0959 (Mische et al., 2007) |
Line # 16.1 | |
| genetic reagent (Drosophila melanogaster) |
UAS-Lis1-RNAi | Graydon Gonsalvez lab (Augusta University); doi:10.1534/genetics. 115.180018 (Liu et al., 2015) |
||
| genetic reagent (Drosophila melanogaster) |
UAS-BicD-RNAi | Bloomington Drosophila Stock Center |
BDSC: 35405; FlyBase ID: FBti0144407; TRiP.GL00325 |
|
| genetic reagent (Drosophila melanogaster) |
UAS-Spindly-RNAi | Bloomington Drosophila Stock Center |
BDSC: 34933; FlyBase ID: FBti0144908; TRiP.HMS01283 |
|
| genetic reagent (Drosophila melanogaster) |
UAS-hook-RNAi | Bloomington Drosophila Stock Center |
BDSC: 64663; FlyBase ID: FBti0183911; TRiP.HMC05698 |
|
| genetic reagent (Drosophila melanogaster) |
UAS-GFP-RNAi | Bloomington Drosophila Stock Center |
BDSC: 41551; FlyBase ID: FBti0148872 |
|
| genetic reagent (Drosophila melanogaster) |
UASp-tdMaple3- αtub84B |
Our lab; doi:10.1073/pnas. 1522424113 (Lu et al., 2016) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-EMTB- 3XTagRFP |
Our lab; doi:10.7554/eLife. 54216 (Lu et al., 2020) |
||
| genetic reagent (Drosophila melanogaster) |
mat αtub67C- EMTB-3XGFP- sqh 3'UTR |
Yu-Chiun Wang lab (RIKEN Center for Biosystems Dynamics Research) (an unpublished gift) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-GFP- Patronin |
Uri Abdu lab (Ben Gurion University); doi:10.7554/eLife.54216 (Lu et al., 2020); doi:10.1083/jcb.201709174 (Lu et al., 2018); doi:10.1242/bio.039552 (Baskar et al., 2019); doi:10.1016 /j.cub. 2021.05.010 (Lu et al., 2021) |
||
| genetic reagent (Drosophila melanogaster) |
Jupiter-GFP | Yale GFP Protein Trap Database doi: 10.1073/pnas. 261408198 (Morin et al., 2001); doi:10.1016 /j.cub. 2013.04.050 (Lu et al., 2013b); doi:10.1091/mbc. E14-10-1423 (Lu et al., 2015); doi:10.1002 /cm. 20124 (Karpova et al., 2006) |
ZCL2183; FlyBase ID: FBal0286259 |
|
| genetic reagent (Drosophila melanogaster) |
UASp-LifeAct- TagRFP |
Bloomington Drosophila Stock Center |
BDSC: 58714; FlyBase ID: FBti0164964 |
|
| genetic reagent (Drosophila melanogaster) |
UASp-F-Tractin- tdTomato |
Bloomington Drosophila stock center; doi:10.1016 /j. ydbio.2014.06.022 (Spracklen et al., 2014) |
BDSC: 58989; FlyBase ID: FBti0164816 |
|
| genetic reagent (Drosophila melanogaster) |
UAS-zip-RNAi | Bloomington Drosophila Stock Center |
BDSC: 37480; FlyBase ID: FBti0146538; TRiP.GL00623 |
|
| genetic reagent (Drosophila melanogaster) |
hs-Flp12; Sco/CyO | Bloomington Drosophila stock center |
BDSC: 1929; FlyBase ID: FBti0000784 |
|
| genetic reagent (Drosophila melanogaster) |
FRTG13 zip2/CyO | Bloomington Drosophila stock center |
BDSC: 8739; FlyBase ID: FBti0001247 and FBal0018863 |
|
| genetic reagent (Drosophila melanogaster) |
FRTG13 Ubi-GFP.nls | Bloomington Drosophila stock center |
BDSC: 5826; FlyBase ID: FBti0001247, FBti0016099 and FBti0016100 |
|
| genetic reagent (Drosophila melanogaster) |
Sqh-GFP | Bloomington Drosophila stock center |
BDSC: 57145; FlyBase ID: FBti0150058 |
|
| genetic reagent (Drosophila melanogaster) |
UASp-Mito- MoxMaple3 |
Our lab; doi:10.1016 / j.cub.2021.05.010 (Lu et al., 2021) |
||
| genetic reagent (Drosophila melanogaster) |
ubi-GFP-Pav | David Glover lab (Caltech); doi: 10.1242/jcs. 115.4.725 (Minestrini et al., 2002) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-RFP-Golgi | Bllomington stock center; doi:10.1038 /s41598- 017-05679-1 (Chowdhary et al., 2017); doi:10.1016 /j.cub. 2021.05.010 (Lu et al., 2021) |
BDSC: 30908; FlyBase ID: FBti0129915 |
|
| genetic reagent (Drosophila melanogaster) |
pDlic-Dlic-GFP | Thomas Hays (University of Minnesota) (Neisch et al., 2021)doi: https://doi.org/10.1101/2021.09.27.462070 |
||
| genetic reagent (Drosophila melanogaster) |
shot∆EGC | Ferenc Jankovics, Institute of Genetics, Biological Research Centre of the Hungarian Academy of Sciences; doi:10.1242/jcs.193003 (Takács et al., 2017) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-shot-RNAi | Our lab; doi:10.1016 / j.cub.2021.05.010 (Lu et al., 2021) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-Dlic-RNAi | Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-GEM | Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-F-Tractin- Myc-BicD |
Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-F-Tractin- GFP-Kin14VIb |
Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-Myc-HA- DlicNT-LOV2 |
Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
||
| genetic reagent (Drosophila melanogaster) |
UASp-Zdk1-DlicCT- sfGFP-Myc |
Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
||
| cell line (Drosophila melanogaster) |
S2R + cells |
Drosophila Genomics Resource Center (DGRC) |
Stock Number: 150; RRID:CVCL_Z831 |
Tested negative for mycoplasma; authenticated by the vendor |
| cell line (Homo sapiens) |
HEK293T cells | ATCC; doi: 10.1096/ fj.201800604R (Robert et al., 2019) |
CRL-3216 | Tested negative for mycoplasma; authenticated by the vendor |
| transfected construct (Drosophila melanogaster) |
pcDNA3.1(+)-Myc- HA-DlicNT-LOV2 |
Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Contains Drosophila
Dlic residues 1–370 and LOVTRAP probe LOV2[WT] |
For transfection of HEK293T cells |
| transfected construct (Drosophila melanogaster) |
pcDNA3.1(+)-Zdk1- DlicCT-sfGFP-Myc |
Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Contains LOVTRAP probe Zdk1 and Drosophila Dlic residues 371–493 |
For transfection of HEK293T cells |
| antibody | anti-Orb antibody (Mouse monoclonal) |
Developmental Studies Hybridoma Bank (DSHB) |
orb 4H8; RRID: AB_528418 |
(1:5) dilution |
| antibody | FITC-conjugated anti-β tubulin antibody (Mouse monoclonal) |
ProteinTech | Cat#: CL488-66240; RRID: AB_2883292 |
(1:100) dilution |
| antibody | anti-Drosophila dynein heavy chain antibody (Mouse monoclonal) |
Developmental Studies Hybridoma Bank (DSHB) |
2C11-2; RRID: AB_2091523 | (1:50) dilution |
| antibody | anti-Myc antibody (Mouse monoclonal) |
doi: 10.1126/science.1061086 (Karcher et al., 2001) | Purified from Hybridoma cell line MYC 1-9E10.2 (RRID:CVCL_G671) |
(1:100) dilution |
| antibody | anti-BicD antibody (Mouse monoclonal) |
Developmental Studies Hybridoma Bank (DSHB) |
anti-Bicaudal-D 4C2; RRID: AB_528103 |
(1:5) dilution |
| antibody | anti-phospho-myosin light chain 2 (Ser19) (Rabbit polyclonal) |
Cell Signaling | Cat# 3671; RRID: AB_330248 | (1:100) dilution |
| antibody | Fluorescein (FITC) affiniPure anti- Mouse IgG (H + L) (Goat polyclonal) |
Jackson ImmunoResearch |
Cat# 115-095-062; RRID: AB_2338594 | 10 µg/ml |
| antibody | Rhodamine (TRITC) affiniPure anti- Mouse IgG (H + L) (Goat polyclonal) |
Jackson ImmunoResearch |
Cat# 115-025-003; RRID: AB_2338478 | 10 µg/ml |
| antibody | Fluorescein (FITC) affiniPure anti- Rabbit IgG (H + L) (Goat polyclonal) |
Jackson ImmunoResearch |
Cat#111-095-003; RRID: AB_2337972 | 10 µg/ml |
| antibody | HRP affiniPureanti-mouse IgG (H + L) (Goat polyclonal) |
Jackson ImmunoResearch |
Cat# 115-035-003; RRID: AB_10015289 | 10 µg/ml |
| recombinant DNA reagent (Drosophila melanogaster) |
pWalium22- Dlic3’UTR-shRNA |
Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Targeting Dlic 3’UTR 401–421 nt, 5’-AGAAATTTAAC AAAAAAAAAA –3’ |
DNA plasmid for generating transgenic flies |
| recombinant DNA reagent (Pyrococcus furiosus) |
pUASp-GEM | Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Subcloned from pCDNA3.1-pCMV-PfV-GS-Sapphire (Addgene plasmid # 116933; RRID:Addgene_116933) | DNA plasmid for generating transgenic flies |
| recombinant DNA reagent (Drosophila melanogaster) |
pUASp-F-Tractin-Myc-BicD | Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Includes the actin binding domain of rat Inositol trisphosphate 3-kinase A (lTPKA residues 9–40), Myc and Drosophila BicD | DNA plasmid for generating transgenic flies |
| recombinant DNA reagent (Physcomitrella patens) |
pUASp-F-Tractin-GFP-Kin14VIb | Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Includes the actin binding domain of rat Inositol trisphosphate 3-kinase A (lTPKA residues 9–40), GFP and Physcomitrella patens kinesin14VIb | DNA plasmid for generating transgenic flies |
| recombinant DNA reagent (Drosophila melanogaster) |
pUASp-Myc-HA-DlicNT-LOV2-attB | Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Contains Drosophila Dlic residues 1–370 and LOVTRAP probe LOV2[WT] | DNA plasmid for generating transgenic flies |
| recombinant DNA reagent (Drosophila melanogaster) |
pUASp-Zdk1-DlicCT-sfGFP-Myc-attB | Generated in this study (available upon request, contact vgelfand@northwestern.edu) |
Contains LOVTRAP probe Zdk1 and Drosophila Dlic residues 371–493 | DNA plasmid for generating transgenic flies |
| chemical compound, drug |
Rhodamine-labeled phalloidin | Thermo Fisher Scientific | Cat# R415 | 0.2 µg/ml |
| chemical compound, drug |
DAPI | Sigma-Aldrich (MilliporeSigma) |
D9542-1MG | 1 µg/ml |
| chemical compound, drug |
16% Paraformaldehyde Aqueous Solution, EM Grade | Fisher Scientific (Electron Microscopy Science) |
Cat# 50-980-487 (EMS 15710) | 4% or 8% for fixation |