Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| cell line (Dictyostelium) | control wild type | Provided by Dr. Günther Gerisch (MPI Martinsried) | Ax2 | Available at Dictybase.org |
| cell line (Dictyostelium) | control wild type | Provided by Dr. Rick Firtel (UCSD) | Ax3 | Available at Dictybase.org |
| cell line (Dictyostelium) | myo7 null | Titus Lab Tuxworth et al., 2001 |
myo7 null HTD17-1 (G1-20) |
Ax3 background |
| cell line (Dictyostelium) | control non-homologous recombinant - myo7 null | Titus Lab Tuxworth et al., 2001 |
G1-21 | Ax3 background |
| cell line (Dictyostelium) | vasp null | Firtel Lab Han et al., 2002 |
vasp - | Ax3 background |
| cell line (Dictyostelium) | dDia2 null | Provided by Dr. Jan Faix (Hannover Med Sch, Hannover Germany) Schirenbeck et al., 2005 | dia - | Ax2 background |
| Other | HL5 Medium including Glucose | Formedium | HLG0103 | Dictyostelium growth medium |
| Database | Dictybase.org | Fey et al., 2013 | ||
| Software | Prism 8.0 | Graphpad | Statistical Analysis | graph preparation, statistical analysis |
| Software | Seven | Petersen et al., 2016 | https://github.com/tituslabumn/Seven | |
| Antibody | Rabbit anti-DdMyo7 | UMN-87 Tuxworth et al., 2005 | 1:2000 | |
| Antibody | mouse monoclonal anti-GFP, clone B34 | Biolegend | 902602 | 1:5000 |
| Antibody | Rabbit polyclonal anti-vasp | Provided by Dr. Jan Faix (Hannover Med Sch, Hannover Germany) Breitsprecher et al., 2008 | 1:500 | |
| Antibody | Rabbit polyclonal anti myoB | Titus Lab Novak et al., 1995 | 1:2000 | |
| Antibody | Rabbit polyclonal anti-mCherry | Proteintech | 26765–1-AP | 1:500 |
| Antibody | Goat anti rabbit secondary | Licor | IR680 | 1:2500-1:10,000 |
| Antibody | Goat anti mouse secondary | Licor | IR800 | 1:2500-1:10,000 |
| Antibody | Mouse monoclonal anti- Dictyostelium actin | Provided by Dr. Günther Gerisch (MPI Martinsried) Westphal et al., 1997 | 1:4000 | |
| Chemical compound | G418 | Fisher Scientific | BP-673 | antibiotic |
| Chemical compound | G418 | Gold Biotechnology | G-418 | antibiotic |
| Chemical compound | Hygromycin B | Gold Biotechnology | H-270 | antibiotic |
| Chemical compound | Doxycycline Hyclate | Sigma | D9891 | antibiotic |
| Chemical compound | Pencillin G Sodium Salt | Sigma | P3032 | antibiotic |
| Chemical compound | Streptomycin Sulfate | Sigma | S9137 | antibiotic |
| Chemical compound, drug | Cytochalasin A | Sigma | C-6637 | |
| Chemical compound, drug | Jasplakinolide | Sigma | J4580 | |
| Chemical compound, drug | Latrunculin A | Sigma | L5163 | |
| Chemical compound, drug | LY294002 | EMD Millipore | 440204 | |
| Chemical compound, drug | Nocodazole | Sigma | M1404 | |
| Chemical compound, drug | Wortmannin | Sigma | W1628 | |
| Chemical compound, drug | CK666 | Gift from Dr. Brad Nolen (U. Oregon) Nolen et al., 2009 | ||
| Chemical compound, drug | Alexa Fluor 568 Phalloidin | Invitrogen | A12380 | |
| Chemical compound, drug | Alexa Fluor 647 Phalloidin | Invitrogen | A22287 | |
| Chemical compound, drug | FM 4-64 Dye | ThermoFisher | T13320 | |
| other | Q5 polymerase | New England Biolabs | M049L | |
| commercial assay or kit | NEBuilder HiFi DNA Assembly | New England Biolabs | E5520S | |
| Recombinant DNA reagent | synthesized gene - mScarlet I | Genscript | actagtggtggttcaggaGTTTCAAAAGGTGAAGCCGTTATTAAAGAATTTATGAGATTCAAGGTTCACATGGAAGGAAGTATGAACGGTCATGAATTTGAGATTGAAGGAGAAGGTGAAGGTAGACCATATGAAGGCACCCAAACAGCTAAATTAAAAGTAACTAAAGGTGGTCCATTACCATTTAGTTGGGATATTTTATCTCCACAATTTATGTATGGTTCACGTGCTTTCAttAAACATCCAGCAGATATTCCAGATTATTATAAACAATCATTTCCAGAA GGTTTTAAATGGGAACGTGTCATGAACTTTGAAGATGGTGGAGCAGTTACAGTCACACAAGATACCTCATTAGAAGATGGTACATTAATATATAAAGTTAAATTACGTGGTACTAATTTTCCACCAGACGGTCCAGTAATGCAAAAAAAAACAATGGGCTGGGAAGCTAGT ACAGAACGTTTATATCCTGAAGATGGTGTCCTTAAAGGCGATATAAAAATGGCCTTGAGATTAAAGGATGGTGGTAGGTATTTAGCAGATTTCAAAACCACTTATAAAGCAAAAAAACCAGTTCAAATGCCAGGTGCATATAATGTTGATAGAAAACTTGATATTACCAGTCATAATGAAGATTACACAGTTGTCGAACAATACGAACGTTCTGAAGGTCGTCATAGCACTGGTGGTATGGATGAATTATACAAATAAgctagc | |
| Recombinant DNA reagent | synthesized gene - mNeon | Genscript | ggatccATGGTGAGTAAAGGTGAAGAAGATAATATGGCATCGTTACCAGCTACACATGAGTTACATATATTCGGTAGCATTAATGGTGTTGATTTTGATATGGTGGGACAAGGTACCGGTAATCCTAATGATGGTTACGAAGAACTAAATTTAAAATCGACTAAAGGTGACTTACAATTTTCTCCATGGATTTTAGTGCCACATATAGGGTATGGTTTTCATCAATACTTACCATATCCAGATGGTATGTCACCATTTCAAGCTGCAATGGTTGATGGATCAGGTTATCAAGTTCATAGAACAATGCAATTTGAAGATGGTGCTTCATTAACTGTTAATTATAGATACACATATGAAGGCTCACATATTAAAGGTGAAGCTCAAGTTAAAGGTACTGGTTTCCCAGCCGATGGCCCAGTTATGACAAATAGTTTAACAGCAGCAGATTGGTGTAGATCCAAAAAAACTTATCCAAATGATAAAACAATTATTTCAACTTTTAAATGGTCATATACAACCGGTAATGGTAAACGTTATCGTTCAACAGCCCGTACAACATATACTTTTGCTAAACCAATGGCAGCTAATTATTTAAAAAATCAACCAATGTATGTTTTTCGTAAAACAGAGTTAAAACATTCAAAAACAGAACTTAATTTTAAAGAATGGCAAAAAGCATTTACAGACGTTATGGGTATGGATGAACTTTATAAGagatct | |
| Sequence-based reagent | CAAX F | IDTDNA | PCR Primer (pDTi346 plasmid) | ttattaTAAAAAAATTAAAATAAAATAAAATCTCGTG |
| Sequence-based reagent | CAAX R | IDTDNA | PCR Primer (pDTi346 plasmid) | tgtacaTTGAGAAGAATAAAATTGATAAACTG |
| Sequence-based reagent | E386V F | IDTDNA | PCR Primer (pDTi364 plasmid) | ttttgtAAATTTTAAAAAGAATAGTTTTGAACAATTTTG |
| Sequence-based reagent | E386V R | IDTDNA | PCR Primer (pDTi364 plasmid) | ccaaagATATCCAATACACCAATAAATGTTG |
| Sequence-based reagent | I426A F | IDTDNA | PCR Primer (pDTi435 plasmid) | AAAAGAAAAAgctAATTGGAGTAAGATCGTATATAATG |
| Sequence-based reagent | I426A R | IDTDNA | PCR Primer (pDTi435 plasmid) | TCATATTCTTCTTGTTCTAATTTAAAAATATG |
| Sequence-based reagent | myi42 | in-house synthesis | PCR Primer (pDTi289 + 2 plasmid) | catgccatggcagcagcagcaACCTTAAAGAGAAAAGCACCAGTCG |
| Sequence-based reagent | myi185 + 2 | IDTDNA | PCR primer (pDTi289 + 2 plasmid) | gctagcaaTTGAGAAGAATAAAATTGATAAACTGAAGC |
| Sequence-based reagent | VASP339* F | IDTDNA | related to pVASP29 plasmid | taataaAGAGCATCTCAACATTAACTAG |
| Sequence-based reagent | VASP339* R | IDTDNA | PCR Primer (pVASP29 plasmid) | cccgggAGCTGATAAGGATGGTGAAG |
| Sequence-based reagent | FAB K-E F | IDTDNA | PCR Primer (pVASP34 plasmid) | gaaatggagGCAGCAGCATCTCAACCAA |
| Sequence-based reagent | FAB K-E R | IDTDNA | PCR primer (pVASP34 plasmid) | ggcttcctcGGCCATAACTTCGGCCAT- |
| Sequence-based reagent | forH4L F | IDTDNA | PCR primer (dDia2-SC plasmid) | AATTGACCAGATCTAATTTGAG |
| Sequence-based reagent | forH8 R | IDTDNA | PCR primer (dDia2 SC plasmid) | actagtTTATTTTTTTAATTGGCCTGATGG |
| Sequence-based reagent | forH9 F | IDTDNA | PCR primer (dDia2-CA SC plasmid) | ggatccATGTCTTTTGATTTAGAGAGTAATAGTAGTGG |
| Sequence-based reagent | forH10 R | IDTDNA | PCR primer (dDia2-CA SC plasmid) | ATTCAAAGATagaagaGTTGGTGATTCTGTCATTG |
| Sequence-based reagent | V1 F | IDTDNA | PCR primer (V-1 SC plasmid) | agatctATGGAAGAACAAAATGATTTCAC |
| Sequence-based reagent | V1 R | IDTDNA | PCR primer (V-1 SC plasmid) | actagtTTATTTTAATAATGCTTTAATATCAGC |
| Transformed construct (Dictyostelium) | GFP-DdMyo7-Tail | Titus Lab Tuxworth et al., 2005 | pDTi35 | pSmall, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | GFP-DdMyo7 | Titus Lab Tuxworth et al., 2001 | pDTi74 | pBS, G418, Integrating |
| Transformed construct (Dictyostelium) | GFP-DdMyo7motor-FD | Titus Lab Arthur et al., 2019 | pDTi490 | pDXA, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | GFP-DdMyo7-KKAA | Titus Lab Arthur et al., 2019 | pDTi321 | pBS, G418, Integrating |
| Transformed construct (Dictyostelium) | GFP-DdMyo7-E386V | Titus Lab this paper | pDTi364 | pBS, G418, Integrating |
| Transformed construct (Dictyostelium) | GFP-DdMyo7-E386V; KKAA | Titus Lab this paper | pDTi386 | pBS, G418, Integrating |
| Transformed construct (Dictyostelium) | GFP-DdMyo7-CAAX | Titus Lab this paper | pDTi346 | pBS, G418, Integrating |
| Transformed construct (Dictyostelium) | GFP-DdMyo7-I426A | Titus Lab this paper | pDTi435 | pTX-GFP, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | GFP-DdMyo7 | Titus Lab this paper | pDTi504 | pDM317, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | mNeon-DdMyo7 | Titus Lab, this paper | pDTi516 | pDM304, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | mNeon-DdMyo7-E386V | Titus Lab, this paper | pDTi527 | pDM304, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | DdMyo7-mCherry | Titus Lab, this paper | pDTi340 | pDM358, Hyg, Extrachromosomal |
| Transformed construct (Dictyostelium) | mApple-DdMyo7 | Titus Lab | pDTi512 | pDM358, Hyg, Extrachromosomal |
| Transformed construct (Dictyostelium) | GFP-VASP | Provided by Dr. Rick Firtel (UCSD) Han et al., 2002 | GFP-VASP | EXP4+, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | VASP | Titus Lab, this paper | VASP | pDM358, Hyg, Extrachromosomal |
| Transformed construct (Dictyostelium) | VASP-∆TET | Titus Lab, this paper | VASP-∆TET | pDM358, Hyg, Extrachromosomal |
| Transformed construct (Dictyostelium) | VASP-FAB | Titus Lab, this paper | VASP-FAB | pDM358, Hyg, Extrachromosomal |
| Transformed construct (Dictyostelium) | diaWT | Provided by Dr. Jan Faix (Hannover Med Sch, Hannover Germany) Schirenbeck et al., 2005 | GFP-dDia2 | pDGFP-MCS, G418, Integrating |
| Transformed construct (Dictyostelium) | diaCA | Titus Lab, this paper | mRFP-mars-diaCA | pDM449, Hyg, Extrachromosomal |
| Transformed construct (Dictyostelium) | GFP-V1 | Titus Lab, this paper | Tetracycline -Inducible GFP-V1 | pDM334, G418, Extrachromosomal |
| Transformed construct (Dictyostelium) | RFP-Lifeact | Brzeska et al., 2014 | pDM358, Hyg, Extrachromosomal | |
| Transformed construct (Dictyostelium) | RFP-LimEΔcoil | Gerisch et al., 2004 | ||
| Transformed construct (Dictyostelium) | GFP-tubulin | Provided by Dr. Günther Gerisch (MPI Martinsried) Neujahr et al., 1998 | pDEXRH, G418, Integrating |