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Structure paper

TitleCryo-EM of α-tubulin isotype-containing microtubules revealed a contracted structure of α4A/β2A microtubules.
Journal, issue, pagesActa Biochim Biophys Sin (Shanghai), Vol. 55, Issue 10, Page 1551-1560, Year 2023
Publish dateJul 13, 2023
AuthorsLei Diao / Wei Zheng / Qiaoyu Zhao / Mingyi Liu / Zhenglin Fu / Xu Zhang / Lan Bao / Yao Cong /
PubMed AbstractMicrotubules are hollow α/β-tubulin heterodimeric polymers that play critical roles in cells. In vertebrates, both α- and β-tubulins have multiple isotypes encoded by different genes, which are ...Microtubules are hollow α/β-tubulin heterodimeric polymers that play critical roles in cells. In vertebrates, both α- and β-tubulins have multiple isotypes encoded by different genes, which are intrinsic factors in regulating microtubule functions. However, the structures of microtubules composed of different tubulin isotypes, especially α-tubulin isotypes, remain largely unknown. Here, we purify recombinant tubulin heterodimers composed of different mouse α-tubulin isotypes, including α1A, α1C and α4A, with the β-tubulin isotype β2A. We further assemble and determine the cryo-electron microscopy (cryo-EM) structures of α1A/β2A, α1C/β2A, and α4A/β2A microtubules. Our structural analysis demonstrates that α4A/β2A microtubules exhibit longitudinal contraction between tubulin interdimers compared with α1A/β2A and α1C/β2A microtubules. Collectively, our findings reveal that α-tubulin isotype composition can tune microtubule structures, and also provide evidence for the "tubulin code" hypothesis.
External linksActa Biochim Biophys Sin (Shanghai) / PubMed:37439022 / PubMed Central
MethodsEM (single particle)
Resolution4.2 - 4.4 Å
Structure data

35790

8ixa

8ixb

EMDB-35790: GMPCPP-Alpha1A/Beta2A-microtubule decorated with kinesin
PDB-8ixa: GMPCPP-Alpha1A/Beta2A-microtubule decorated with kinesin non-seam region
PDB-8ixb: GMPCPP-Alpha1A/Beta2A-microtubule decorated with kinesin seam region
Method: EM (single particle) / Resolution: 4.2 Å

35791

8ixd

8ixe

EMDB-35791: GMPCPP-Alpha1C/Beta2A-microtubule decorated with kinesin
PDB-8ixd: GMPCPP-Alpha1C/Beta2A-microtubule decorated with kinesin non-seam region
PDB-8ixe: GMPCPP-Alpha1C/Beta2A-microtubule decorated with kinesin seam region
Method: EM (single particle) / Resolution: 4.4 Å

35792

8ixf

8ixg

EMDB-35792: GMPCPP-Alpha4A/Beta2A-microtubule decorated with kinesin
PDB-8ixf: GMPCPP-Alpha4A/Beta2A-microtubule decorated with kinesin non-seam region
PDB-8ixg: GMPCPP-Alpha4A/Beta2A-microtubule decorated with kinesin seam region
Method: EM (single particle) / Resolution: 4.4 Å

Chemicals

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

ChemComp-G2P:
PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER / GMP-CPP, energy-carrying molecule analogue*YM

ChemComp-ATP:
ADENOSINE-5'-TRIPHOSPHATE / ATP, energy-carrying molecule*YM / Adenosine triphosphate

Source
  • mus musculus (house mouse)
  • homo sapiens (human)
KeywordsSTRUCTURAL PROTEIN / microtubule / tubulin isotype / cryo-EM structure

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