MassIVEapplication/xmlftp://massive-ftp.ucsd.edu/v05/MSV000090661/primary200OK0000ProteomicsKent HillJames WohlschlegelOrbitrap Fusion LumosTrypanosoma Brucei (ncbitaxon:5691)https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=f65ea18697f14036a054023223b03d2bUCLAjwohl@ucla.edukenthill@microbio.ucla.eduMassIVE31MS:1002864 - No post-translational-modifications are included in the identified peptides of this datasetMotility of pathogenic protozoa depends on flagella (synonymous with cilia) with axonemes containing nine doublet microtubules (DMTs) and two singlet microtubules. Microtubule inner proteins (MIPs) within DMTs influence axoneme stability and motility and provide lineage-specific adaptations, but individual MIP functions and assembly mechanisms are mostly unknown. Here, we show in the sleeping sickness parasite Trypanosoma brucei, that FAP106, a conserved MIP at the DMT inner junction, is required for trypanosome motility and functions as a critical interaction hub, directing assembly of several conserved and lineage-specific MIPs. We use comparative cryogenic electron tomography (cryoET) and quantitative proteomics to identify MIP candidates. Using RNAi knockdown together with fitting of AlphaFold models into cryoET maps, we demonstrate that one of these candidates, MC8, is a trypanosome-specific MIP required for parasite motility. Our work advances understanding of MIP assembly mechanisms and identifies lineage-specific motility proteins that are attractive targets to consider for therapeutic intervention.FAP106 is an interaction hub for assembling microtubule inner proteins at the cilium inner junction.Shimogawa Michelle M MM, Wijono Angeline S AS, Wang Hui H, Zhang Jiayan J, Sha Jihui J, Szombathy Natasha N, Vadakkan Sabeeca S, Pelayo Paula P, Jonnalagadda Keya K, Wohlschlegel James J, Zhou Z Hong ZH, Hill Kent L KLjunction, boundary, Nervous system-specific RNA-binding protein Hel-N1, HELN1, Motile, xel-1, HEL-N1, Motile Cilia, Proteins, Nodal Cilium, primary cilium, Hub, HUB, Gene, Cilium, flagellum, Primary, elrB, neurotubule, elavl2, Nodal Cilia, Hu-antigen B, Nervous system-specific RNA-binding protein Mel-N1, Cilia, Protein Gene Products, Gene Proteins, Primary Cilia, el-1, Xel-1, breakpoint., anatomical junction, Protein, Primary Cilium, p45, Gene Products, Nodal, HuB, hub, microtubuli, Motile Cilium, hel-n1, microtubule-based flagellum, ELAV-like neuronal protein 1, heln1, Microtubule, microtubulus, mel-N1, eukaryotic flagellumjunction, Nervous system-specific RNA-binding protein Hel-N1, HEL-N1, Proteins, primary cilium, Gene, Cilium, flagellum, Primary, elavl2, Nodal Cilia, Nervous system-specific RNA-binding protein Mel-N1, Primary Cilia, TPSG1, el-1, S-adenosyl-L-methionine:thiol S-methyltransferase activity, anatomical junction, Protein, Gene Products, HuB, hel-n1, ELAV-like neuronal protein 1, heln1, eukaryotic flagellum, boundary, HELN1, Motile, xel-1, Motile Cilia, Nodal Cilium, Hub, HUB, elrB, Hu-antigen B, Cilia, Protein Gene Products, Gene Proteins, Xel-1, breakpoint., PRSS31, Primary Cilium, p45, Nodal, hub, Motile Cilium, TMT, microtubule-based flagellum, thiol methyltransferase activity, mel-N1post-transcriptional gene silencing by RNA, rasGAP, Trypanosoma brucei subgroup, IPO2, Peptidomics, Kpnb2, e(-), number, Cilium, Gene, Trypanosoma (Trypanozoon) brucei, neurotubule, electron, GAP1, elavl2, presence, African trypanosomiasis, Nervous system-specific RNA-binding protein Mel-N1, Intervention or Procedure, Primary Cilia, Trypanosoma, Tomographies, Readability, gap1, CG8374, Gene Products, Post Transcriptional, MIP, hel-n1, microtubulus, heln1, KPNB2, AU021749, outer-doublet microtubules, l(2)02695, 0481/03, flagellar axoneme, interventionDescription, EPB49, Elektron, Nodal Cilium, Gap 1, beta-particle, Interventional, mip, MIP1, quelling, Cosuppression, Fast, CG4539, Gene Silencings, M9 region interaction protein, e, D13Ertd688e, e-, SURGICAL AND MEDICAL PROCEDURES., mitochondrial intermediate precursor-processing proteinase activity, bruceus, breakpoint, Nodal, CG6721, Axonemes, microtubuli, DmelCG8374, flagellum axoneme, HAT, sleeping sickness, Electrons, Positron, junction, Intervention Strategies, Nervous system-specific RNA-binding protein Hel-N1, RNA, Flagellum, Trypanosoma brucei bruceus, African sleeping sickness, ciliary axoneme, HEL-N1, Negatron, Proteins, TRN, posttranscriptional gene silencing by siRNA, helminthology, Maps, Negatrons, Primary, Positrons, Procedure, cosuppression, Nodal Cilia, RNAi, el-1, count in organism, 5730405E07Rik, Interference, anatomical junction, African Trypanosomiasis, Protein, sxt, cilium axoneme, Karyopherin beta-2, Fast Electron, HuB, Africam sleeping sickness, rI533, Gap, GAP, Fast Electrons, RNA Silencing, human African trypanosomiasis, brucei, Microtubule, ELAV-like neuronal protein 1, Silencing, Post-Transcriptional Gene, s048103, boundary, Post-Transcriptional Gene Silencings, Intervention, Ras-GAP, HELN1, Motile, DmelCG6721, xel-1, negatron, anon-WO0118547.317, Posttranscriptional, HMIP, Posttranscriptional Gene Silencing, Motile Cilia, Trypanosoma bruceus, Hub, HUB, gap, Electron, Importin beta-2, elrB, Understanding, protozoology, Co Suppression, Trypanosoma brucei, beta, Post Transcriptional Gene Silencing, Cilia, Hu-antigen B, beta(-), Protein Gene Products, Gene Proteins, BcDNA:RE14949, Xel-1, Post-Transcriptional, Gene Silencing, Parasite, DmelCG4539, DMT, Dmt, brucei brucei, protozoa, Primary Cilium, 3.4.24.59, p45, hub, Motile Cilium, Co-Suppression, parasites, Post-Transcriptional Gene Silencing, mel-N1rasGAP, Flagellum, IPO2, determination, Kpnb2, TRN, primary cilium, Skeletons, helminthology, flagellum, GAP1, protozoa., TPSG1, S-adenosyl-L-methionine:thiol S-methyltransferase activity, 5730405E07Rik, gap1, chemical analysis, sxt, Karyopherin beta-2, MIP, rI533, Gap, GAP, eukaryotic flagellum, KPNB2, AU021749, Ras-GAP, l(2)02695, DmelCG6721, HMIP, Gap 1, mip, gap, Importin beta-2, MIP1, protozoology, CG4539, BcDNA:RE14949, M9 region interaction protein, D13Ertd688e, Parasite, DmelCG4539, PRSS31, mitochondrial intermediate precursor-processing proteinase activity, 3.4.24.59, CG6721, assay, parasites, TMT, microtubule-based flagellum, thiol methyltransferase activity0falseTMT analysis of flagellum skeletons from MIP knockdown parasitesTMT data associated with Shimogawa et al. 'FAP106 is an interaction hub required for stable assembly of conserved and lineage-specific proteins at the cilium inner junction'Mon Nov 07 20:11:00 GMT 2022MSV00009066137633952