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TitleSPACA9 is a lumenal protein of human ciliary singlet and doublet microtubules.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 119, Issue 41, Page e2207605119, Year 2022
Publish dateOct 11, 2022
AuthorsMiao Gui / Jacob T Croft / Davide Zabeo / Vajradhar Acharya / Justin M Kollman / Thomas Burgoyne / Johanna L Höög / Alan Brown /
PubMed AbstractThe cilium-centrosome complex contains triplet, doublet, and singlet microtubules. The lumenal surfaces of each microtubule within this diverse array are decorated by microtubule inner proteins ...The cilium-centrosome complex contains triplet, doublet, and singlet microtubules. The lumenal surfaces of each microtubule within this diverse array are decorated by microtubule inner proteins (MIPs). Here, we used single-particle cryo-electron microscopy methods to build atomic models of two types of human ciliary microtubule: the doublet microtubules of multiciliated respiratory cells and the distal singlet microtubules of monoflagellated human spermatozoa. We discover that SPACA9 is a polyspecific MIP capable of binding both microtubule types. SPACA9 forms intralumenal striations in the B tubule of respiratory doublet microtubules and noncontinuous spirals in sperm singlet microtubules. By acquiring new and reanalyzing previous cryo-electron tomography data, we show that SPACA9-like intralumenal striations are common features of different microtubule types in animal cilia. Our structures provide detailed references to help rationalize ciliopathy-causing mutations and position cryo-EM as a tool for the analysis of samples obtained directly from ciliopathy patients.
External linksProc Natl Acad Sci U S A / PubMed:36191189 / PubMed Central
MethodsEM (tomography) / EM (subtomogram averaging) / EM (single particle)
Resolution3.6 - 20.0 Å
Structure data

EMDB-14723: Tomogram of bovine sperm singlet microtubules
Method: EM (tomography)

EMDB-14730: Tomogram of chicken sperm distal microtubules
Method: EM (tomography)

EMDB-14732: Tomogram of frog sperm singlet distal microtubules
Method: EM (tomography)

EMDB-15451: Sub-tomogram average of TAILS in bovine sperm tail tip singlet microtubules.
Method: EM (subtomogram averaging) / Resolution: 20.0 Å

26611

7un1

EMDB-26611: Composite cryo-EM density map of the 8-nm repeat of the human sperm tip singlet microtubule
PDB-7un1: 8-nm repeat of the human sperm tip singlet microtubule
Method: EM (single particle) / Resolution: 6.0 Å

26624

7ung

EMDB-26624: Composite cryo-EM density map of the 48-nm repeat of the human respiratory doublet microtubule
PDB-7ung: 48-nm repeat of the human respiratory doublet microtubule
Method: EM (single particle) / Resolution: 3.6 Å

Chemicals

ChemComp-GDP:
GUANOSINE-5'-DIPHOSPHATE / GDP, energy-carrying molecule*YM / Guanosine diphosphate

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM / Guanosine triphosphate

ChemComp-MG:
Unknown entry

Source
  • Bos taurus (cattle)
  • Gallus gallus (chicken)
  • Xenopus tropicalis (tropical clawed frog)
  • homo sapiens (human)
  • human (human)
KeywordsSTRUCTURAL PROTEIN / cilia / microtubule / sperm / cell motility

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