Computational protocol: An unconventional interaction between Dis1/TOG and Mal3/EB1 in fission yeast promotes the fidelity of chromosome segregation

Similar protocols

Protocol publication

[…] X-ray data were collected on beamlines I04 and I03 of the Diamond Light Source. Data were integrated and scaled using the Xia2 pipeline. The unliganded Mal3 EBH (hereafter Mal3) domain was solved using ab initio molecular replacement as implemented in the ARCIMBOLDO-LIGHT package (). 20-residue α-helical segments were used as a search model. A strong (30.2% origin) native Patterson peak at 0.155, 0.5, 0.5 in the native data suggested the presence of translational pseudo-symmetry, and the appropriate correction was applied in Phaser. The preliminary poly-alanine model was then assigned sequence in Buccaneer, and manually rebuilt and refined with Coot () and Refmac5 (). The final model was refined with anisotropic temperature factors applied to all atoms. The Mal3–Dis1 structure was solved by molecular replacement in Phaser () using the apo-Mal3 structure as a search model. Model building and refinement were carried out as for the apo-structure. Clear density could be seen for the bound Dis1 peptides in an unbiased difference map at both homodimer interfaces and was built in the final stages of refinement. All data collection and refinement statistics are provided in Table S4. […]

Pipeline specifications

Software tools xia2, Buccaneer, Coot, REFMAC5
Applications Small-angle scattering, Protein structure analysis
Organisms Dipturus trachyderma, Schizosaccharomyces pombe