Supplementary Figure 3: The interaction between ASPM and katanin requires a conserved repeat sequence of ASPM and the p60N/p80C heterodimer.
From: Microtubule minus-end regulation at spindle poles by an ASPM–katanin complex
(a) Alignment of katanin-binding linear motifs of ASPM from several vertebrate species. Note the complete conservation of the phenylalanine residue corresponding to F352 in the third repeat of mouse ASPM (red arrowhead below the alignment). (b) Streptavidin pull down assays with extracts of HEK293T cells expressing Biotinylation tagged (Bio)-GFP-tagged wild-type (WT) p80 or its indicated mutants together with GFP-tagged WT p60 or the indicated mutants. None of the analyzed mutations perturbed the p60–p80 interaction. See also Supplementary Fig. 8e. (c) Electron density maps of ASPMp in the p60N/p80C/ASPMp complex structure. Only residues S351, F352 and L353 of the ASPMp peptide (LSPDSFLND, residues 347–355 of mouse ASPM) are visible. The SigmaA-weighted 2mFo-DFc (left) and mFo-DFc (right) omit maps (green mash) are contoured at +1.0σ and +3.0σ, respectively.
