diff --git a/README.md b/README.md index 3e14a2f6161194e0665ec6386d699261fc174a68..44219b8b6d83c516ff09ed0ed03915ce7ad24a0b 100644 --- a/README.md +++ b/README.md @@ -8,7 +8,7 @@ https://www.pnas.org/doi/10.1073/pnas.2404091121 1. [Abstract](## Abstract) 2. [Studies](## Studies) -3. [Assays](## Assays) (https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays?ref_type=heads) +3. [Assays](## Assays) 4. [Licence](## License) ## Abstract @@ -16,70 +16,70 @@ Microtubule-dependent endosomal transport is crucial for polar growth, ensuring ## Studies -The [Studies (S)](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/studies?ref_type=heads) are named after the chapters of the publication. +The [Studies (S)](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/studies?ref_type=heads) are named after the chapters of the publication. -S1_MLLE3 of Rrm4 Constitutes a Different Type of Seven-Helix MLLE Domain -S2_MLLE3 of Rrm4 Recognizes PAM2-Like Sequences with a Defined Binding Pocket -S3_Identifying an Essential FxY Core in PAM2-Like Peptides of Upa1 -S4_MLLE3 of Rrm4 Is Necessary and Sufficient for Endosomal Attachment -S5_Deciphering the Binding Code for MLLE of Pab1 and MLLE3 of Rrm4 -S6_Identification of Unrecognized Rrm4 Interaction Partners -S7_Human MLLE Domains Differentiate between Binding Partners +S1_MLLE3 of Rrm4 Constitutes a Different Type of Seven-Helix MLLE Domain +S2_MLLE3 of Rrm4 Recognizes PAM2-Like Sequences with a Defined Binding Pocket +S3_Identifying an Essential FxY Core in PAM2-Like Peptides of Upa1 +S4_MLLE3 of Rrm4 Is Necessary and Sufficient for Endosomal Attachment +S5_Deciphering the Binding Code for MLLE of Pab1 and MLLE3 of Rrm4 +S6_Identification of Unrecognized Rrm4 Interaction Partners +S7_Human MLLE Domains Differentiate between Binding Partners ## Assays The [Assays](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays?ref_type=heads) folder contain the actual raw/processed data of the experiments in the *dataset* folder and the corresponding protocols in the *protocol* folder. ### Chapter 1: MLLE3 of Rrm4 Constitutes a Different Type of Seven-Helix MLLE Domain -[S1_A1_Protein structure prediction](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A1_Protein%20structure%20prediction?ref_type=heads) Figure 1B, Figure S1F -[S1_A2_Protein Purification for structural studies](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A2_Protein%20Purification%20for%20structural%20studies?ref_type=heads) Figure S1A -[S1_A3_Size exclusion chromatography of H-Rrm4-MLLE3](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A3_Size%20exclusion%20chromatography%20of%20H-Rrm4-MLLE3?ref_type=heads) Figure S1B -[S1_A4_Western blot analysis of GST pull-down experiments with MLLE3 versions](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A4_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20with%20MLLE3%20versions?ref_type=heads) Figure 1C, Figure S1C -[S1_A5_Isothermal Titration Calorimetry_ITC_using Rrm4 MLLE3](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A6_X-ray%20structure%20of%20Rrm4-MLLE3?ref_type=heads) Figure 1D, Figure S1D -[S1_A6_X-ray structure of Rrm4-MLLE3](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A6_X-ray%20structure%20of%20Rrm4-MLLE3?ref_type=heads) Figure 1B, Figure S1F -[S1_A7_Multiple sequence alignment of Rrm4-MLLE3 orthologs of representative fungi from Basidiomycota](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A7_Multiple%20sequence%20alignment%20of%20Rrm4-MLLE3%20orthologs%20of%20representative%20fungi%20from%20Basidiomycota?ref_type=heads) Figure S1F +[S1_A1_Protein structure prediction](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A1_Protein%20structure%20prediction?ref_type=heads) Figure 1B, Figure S1F +[S1_A2_Protein Purification for structural studies](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A2_Protein%20Purification%20for%20structural%20studies?ref_type=heads) Figure S1A +[S1_A3_Size exclusion chromatography of H-Rrm4-MLLE3](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A3_Size%20exclusion%20chromatography%20of%20H-Rrm4-MLLE3?ref_type=heads) Figure S1B +[S1_A4_Western blot analysis of GST pull-down experiments with MLLE3 versions](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A4_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20with%20MLLE3%20versions?ref_type=heads) Figure 1C, Figure S1C +[S1_A5_Isothermal Titration Calorimetry_ITC_using Rrm4 MLLE3](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A6_X-ray%20structure%20of%20Rrm4-MLLE3?ref_type=heads) Figure 1D, Figure S1D +[S1_A6_X-ray structure of Rrm4-MLLE3](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A6_X-ray%20structure%20of%20Rrm4-MLLE3?ref_type=heads) Figure 1B, Figure S1F +[S1_A7_Multiple sequence alignment of Rrm4-MLLE3 orthologs of representative fungi from Basidiomycota](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S1_A7_Multiple%20sequence%20alignment%20of%20Rrm4-MLLE3%20orthologs%20of%20representative%20fungi%20from%20Basidiomycota?ref_type=heads) Figure S1F ### Chapter 2: MLLE3 of Rrm4 Recognizes PAM2-Like Sequences with a Defined Binding Pocket -[S2_A1_Structural analysis of MLLE3 of Rrm4](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S2_A1_Structural%20analysis%20of%20MLLE3%20of%20Rrm4?ref_type=heads) Figure 2A, Figure 2B -[S2_A2_Western blot analysis of GST pull-down experiments with MLLE3 variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S2_A2_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20with%20MLLE3%20variants?ref_type=heads) Figure S2A +[S2_A1_Structural analysis of MLLE3 of Rrm4](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S2_A1_Structural%20analysis%20of%20MLLE3%20of%20Rrm4?ref_type=heads) Figure 2A, Figure 2B +[S2_A2_Western blot analysis of GST pull-down experiments with MLLE3 variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S2_A2_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20with%20MLLE3%20variants?ref_type=heads) Figure S2A ### Chapter 3: Identifying an Essential FxY Core in PAM2-Like Peptides of Upa1 -[S3_A1_Western blot analysis of GST pull-down experiments with GST-MLLE3 and PAM2L variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S3_A1_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20with%20GST-MLLE3%20and%20PAM2L%20variants?ref_type=heads) Figure S2A-C, Figure S2G -[S3_A2_Structural comparison of MLLE3 in U_maydis and R_irregularis](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S3_A2_Structural%20comparison%20of%20MLLE3%20in%20U_maydis%20and%20R_irregularis?ref_type=heads) Figure 2B, Figure S2D-F -[S3_A3_Western blot analyis of Rrm4-MLLE3 from U_maydis and R_irregularis](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S3_A3_Western%20blot%20analyis%20of%20Rrm4-MLLE3%20from%20U_maydis%20and%20R_irregularis?ref_type=heads) Figure S2C, Figure S2G +[S3_A1_Western blot analysis of GST pull-down experiments with GST-MLLE3 and PAM2L variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S3_A1_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20with%20GST-MLLE3%20and%20PAM2L%20variants?ref_type=heads) Figure S2A-C, Figure S2G +[S3_A2_Structural comparison of MLLE3 in U_maydis and R_irregularis](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S3_A2_Structural%20comparison%20of%20MLLE3%20in%20U_maydis%20and%20R_irregularis?ref_type=heads) Figure 2B, Figure S2D-F +[S3_A3_Western blot analyis of Rrm4-MLLE3 from U_maydis and R_irregularis](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S3_A3_Western%20blot%20analyis%20of%20Rrm4-MLLE3%20from%20U_maydis%20and%20R_irregularis?ref_type=heads) Figure S2C, Figure S2G ### Chapter 4: MLLE3 of Rrm4 Is Necessary and Sufficient for Endosomal Attachment -[S4_A1_Hyphal growth of strains expressing Rrm4-Kat with MLLE3 variants in AB33 background](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A1_Hyphal%20growth%20of%20strains%20expressing%20Rrm4-Kat%20with%20MLLE3%20variants%20in%20AB33%20background?ref_type=heads) Figure 3B-C, Figure S3 -[S4_A2_Quantification of hyphal growth of AB33 derivatives expressing Rrm4-MLLE variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A2_Quantification%20of%20hyphal%20growth%20of%20AB33%20derivatives%20expressing%20Rrm4-MLLE%20variants?ref_type=heads) Figure 3C -[S4_A3_Micrographs and Kymograph measurement of Rrm4-Kat_MLLE3 variants movement in hyphae](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A3_Micrographs%20and%20Kymograph%20measurement%20of%20Rrm4-Kat_MLLE3%20variants%20movement%20in%20hyphae?ref_type=heads) Figure 3D, Figure S3B-C -[S4_A4_Quantification of processive Rrm4-Kat signals - velocity and distance travelled](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A4_Quantification%20of%20processive%20Rrm4-Kat%20signals%20-%20velocity%20and%20distance%20travelled?ref_type=heads) Figure S3D-I +[S4_A1_Hyphal growth of strains expressing Rrm4-Kat with MLLE3 variants in AB33 background](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A1_Hyphal%20growth%20of%20strains%20expressing%20Rrm4-Kat%20with%20MLLE3%20variants%20in%20AB33%20background?ref_type=heads) Figure 3B-C, Figure S3 +[S4_A2_Quantification of hyphal growth of AB33 derivatives expressing Rrm4-MLLE variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A2_Quantification%20of%20hyphal%20growth%20of%20AB33%20derivatives%20expressing%20Rrm4-MLLE%20variants?ref_type=heads) Figure 3C +[S4_A3_Micrographs and Kymograph measurement of Rrm4-Kat_MLLE3 variants movement in hyphae](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A3_Micrographs%20and%20Kymograph%20measurement%20of%20Rrm4-Kat_MLLE3%20variants%20movement%20in%20hyphae?ref_type=heads) Figure 3D, Figure S3B-C +[S4_A4_Quantification of processive Rrm4-Kat signals - velocity and distance travelled](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S4_A4_Quantification%20of%20processive%20Rrm4-Kat%20signals%20-%20velocity%20and%20distance%20travelled?ref_type=heads) Figure S3D-I ### Chapter 5: Deciphering the Binding Code for MLLE of Pab1 and MLLE3 of Rrm4 -[S5_A1_Protein structure prediction](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A1_Protein%20structure%20prediction?ref_type=heads) Figure S4A-B -[S5_A2_Purification of H-Pab1-MLLE for structural studies](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A2_Purification%20of%20H-Pab1-MLLE%20for%20structural%20studies?ref_type=heads) Figure S4A -[S5_A3_Size exclusion chromotography of H-Pab1-MLLE](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A3_Size%20exclusion%20chromotography%20of%20H-Pab1-MLLE?ref_type=heads) Figure S4B -[S5_A4_Isothermal Titration calorimetry-ITC with H-Pab1-MLLE](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A4_Isothermal%20Titration%20calorimetry-ITC%20with%20H-Pab1-MLLE?ref_type=heads) Figure S4C -[S5_A5_X-ray structure analysis of H-Pab1-MLLE](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A5_X-ray%20structure%20analysis%20of%20H-Pab1-MLLE?ref_type=heads) Figure S4D -[S5_A6_Western blot analysis of GST pull-down experiments using G-Pab1-MLLE and PAM2 variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A6_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20using%20G-Pab1-MLLE%20and%20PAM2%20variants?ref_type=heads) Figure 4C, Figure 4E -[S5_A7_Multiple sequence alignment of Pab1-MLLE orthologs](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A7_Multiple%20sequence%20alignment%20of%20Pab1-MLLE%20orthologs?ref_type=heads) Figure 4D, Figure S4E +[S5_A1_Protein structure prediction](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A1_Protein%20structure%20prediction?ref_type=heads) Figure S4A-B +[S5_A2_Purification of H-Pab1-MLLE for structural studies](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A2_Purification%20of%20H-Pab1-MLLE%20for%20structural%20studies?ref_type=heads) Figure S4A +[S5_A3_Size exclusion chromotography of H-Pab1-MLLE](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A3_Size%20exclusion%20chromotography%20of%20H-Pab1-MLLE?ref_type=heads) Figure S4B +[S5_A4_Isothermal Titration calorimetry-ITC with H-Pab1-MLLE](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A4_Isothermal%20Titration%20calorimetry-ITC%20with%20H-Pab1-MLLE?ref_type=heads) Figure S4C +[S5_A5_X-ray structure analysis of H-Pab1-MLLE](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A5_X-ray%20structure%20analysis%20of%20H-Pab1-MLLE?ref_type=heads) Figure S4D +[S5_A6_Western blot analysis of GST pull-down experiments using G-Pab1-MLLE and PAM2 variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A6_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20using%20G-Pab1-MLLE%20and%20PAM2%20variants?ref_type=heads) Figure 4C, Figure 4E +[S5_A7_Multiple sequence alignment of Pab1-MLLE orthologs](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S5_A7_Multiple%20sequence%20alignment%20of%20Pab1-MLLE%20orthologs?ref_type=heads) Figure 4D, Figure S4E ### Chapter 6: Identification of Unrecognized Rrm4 Interaction Partners -[S6_A1_Multiple Sequence alignment of vps8 and Taf7 orthologs](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A1_Multiple%20Sequence%20alignment%20of%20vps8%20and%20Taf7%20orthologs?ref_type=heads) Figure 5A, Figure S5A-B) -[S6_A2_Protein structure prediction using Alphafold2](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A2_Protein%20structure%20prediction%20using%20Alphafold2?ref_type=heads) Figure 5B, Figure S5C, Figure S5E -[S6_A3_Western blot analysis of GST pull-down experiments using predicted PAM2L variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A3_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20using%20predicted%20PAM2L%20variants?ref_type=heads) Figure 5C,Figure S5D -[S6_A4_Hyphal growth of strains expressing Rrm4-Kat and Vps8-Gfp variants in AB33 background](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A4_Hyphal%20growth%20of%20strains%20expressing%20Rrm4-Kat%20and%20Vps8-Gfp%20variants%20in%20AB33%20background?ref_type=heads) Figure S6A-C -[S6_A5_Quantification of hyphal growth of AB33 derivatives expressing Rrm4-Kat and Vps8-Gfp variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A5_Quantification%20of%20hyphal%20growth%20of%20AB33%20derivatives%20expressing%20Rrm4-Kat%20and%20Vps8-Gfp%20variants?ref_type=heads) Figure S6 A-B -[S6_A6_Micrographs and Kymograph measurement of Rrm4-Kat movement in Vps8-Gfp variant hypha](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A6_Micrographs%20and%20Kymograph%20measurement%20of%20Rrm4-Kat%20movement%20in%20Vps8-Gfp%20variant%20hypha?ref_type=heads) Figure 5E, Figure S7D -[S6_A7_Quantification of processive Rrm4-Kat signals_velocity and distance travelled in vps8-gfp variant hypha](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A7_Quantification%20of%20processive%20Rrm4-Kat%20signals_velocity%20and%20distance%20travelled%20in%20vps8-gfp%20variant%20hypha?ref_type=heads) Figure S7 -[S6_A8_Quantification of Rrm4-Kat signal accumulated at the tip](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A8_Quantification%20of%20Rrm4-Kat%20signal%20accumulated%20at%20the%20tip?ref_type=heads) Figure S6C +[S6_A1_Multiple Sequence alignment of vps8 and Taf7 orthologs](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A1_Multiple%20Sequence%20alignment%20of%20vps8%20and%20Taf7%20orthologs?ref_type=heads) Figure 5A, Figure S5A-B +[S6_A2_Protein structure prediction using Alphafold2](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A2_Protein%20structure%20prediction%20using%20Alphafold2?ref_type=heads) Figure 5B, Figure S5C, Figure S5E +[S6_A3_Western blot analysis of GST pull-down experiments using predicted PAM2L variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A3_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20using%20predicted%20PAM2L%20variants?ref_type=heads) Figure 5C,Figure S5D +[S6_A4_Hyphal growth of strains expressing Rrm4-Kat and Vps8-Gfp variants in AB33 background](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A4_Hyphal%20growth%20of%20strains%20expressing%20Rrm4-Kat%20and%20Vps8-Gfp%20variants%20in%20AB33%20background?ref_type=heads) Figure S6A-C +[S6_A5_Quantification of hyphal growth of AB33 derivatives expressing Rrm4-Kat and Vps8-Gfp variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A5_Quantification%20of%20hyphal%20growth%20of%20AB33%20derivatives%20expressing%20Rrm4-Kat%20and%20Vps8-Gfp%20variants?ref_type=heads) Figure S6 A-B +[S6_A6_Micrographs and Kymograph measurement of Rrm4-Kat movement in Vps8-Gfp variant hypha](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A6_Micrographs%20and%20Kymograph%20measurement%20of%20Rrm4-Kat%20movement%20in%20Vps8-Gfp%20variant%20hypha?ref_type=heads) Figure 5E, Figure S7D +[S6_A7_Quantification of processive Rrm4-Kat signals_velocity and distance travelled in vps8-gfp variant hypha](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A7_Quantification%20of%20processive%20Rrm4-Kat%20signals_velocity%20and%20distance%20travelled%20in%20vps8-gfp%20variant%20hypha?ref_type=heads) Figure S7 +[S6_A8_Quantification of Rrm4-Kat signal accumulated at the tip](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S6_A8_Quantification%20of%20Rrm4-Kat%20signal%20accumulated%20at%20the%20tip?ref_type=heads) Figure S6C ### Chapter 7: Human MLLE Domains Differentiate between Binding Partners -[S7_A1_Multiple sequence alignment of Mkrn1 orthologs](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S7_A1_Multiple%20sequence%20alignment%20of%20Mkrn1%20orthologs?ref_type=heads) Figure 6A-B, Figure S8A -[S7_A2_Protein structure prediction of MKRN1 using Alphafold2](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S7_A2_Protein%20structure%20prediction%20of%20MKRN1%20using%20Alphafold2?ref_type=heads) Figure 6C -[S7_A3_Western blot analysis of GST pull-down experiments using human PAM2L variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S7_A3_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20using%20human%20PAM2L%20variants?ref_type=heads) Figure 6D, Figure S8B +[S7_A1_Multiple sequence alignment of Mkrn1 orthologs](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S7_A1_Multiple%20sequence%20alignment%20of%20Mkrn1%20orthologs?ref_type=heads) Figure 6A-B, Figure S8A +[S7_A2_Protein structure prediction of MKRN1 using Alphafold2](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S7_A2_Protein%20structure%20prediction%20of%20MKRN1%20using%20Alphafold2?ref_type=heads) Figure 6C +[S7_A3_Western blot analysis of GST pull-down experiments using human PAM2L variants](https://git.nfdi4plants.org/devan/2024_Devan-Sai_et_al_PNAS/-/tree/main/assays/S7_A3_Western%20blot%20analysis%20of%20GST%20pull-down%20experiments%20using%20human%20PAM2L%20variants?ref_type=heads) Figure 6D, Figure S8B ## License Copyright © 2024 the Author(s). 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