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. 2021 Oct 28;49(20):11653–11665. doi: 10.1093/nar/gkab942

Figure 1.

Figure 1.

CST DNA-binding mutants maintain subunit assembly and mostly maintain pol α-primase binding. (A) Location of mutated amino acids relative to the DNA (half opaque surface representation, orange) in the cryo-EM structure of CST (22). Grey ribbon, CTC1. Dark surface, STN1. (B) All mutants maintain assembly of CTC1, STN1 and TEN1 subunits. Insect cell recombinant CST (shown here on WT CST gel) was always included to allow plotting a standard curve to calculate concentration of HEK cell CST. Lack of reliable anti-TEN1 antibody led us to probe the HA-tagged TEN1 with anti-HA antibody; the insect cell TEN1 lacked this tag, so was not revealed. The lower band of CTC1, of unknown origin, was consistently missing in the g1.1 mutant. (C) WT CST and all mutants except g1.1 co-purify with pol α-primase, shown here for the two pol α subunits and in Supplementary Figure S1 for the primase subunits. In panels (B) and (C), wedges indicate successive two-fold dilutions of protein.