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. Author manuscript; available in PMC: 2016 Mar 3.
Published in final edited form as: Cell Rep. 2015 Feb 26;10(8):1288–1296. doi: 10.1016/j.celrep.2015.01.054

Figure 4. C6orf211 codes for Armt1 - a carboxyl methyltransferase capable of modifying PCNA.

Figure 4

(A) Recombinant His-tagged C6orf211 was expressed in insect cells, isolated by Ni2+ Sepharose chromatography prior to 10% SDS-PAGE and colloidal Coomassie blue staining. (B) C6orf211 (0.2μg) was assayed by vapor diffusion in absence and presence of PCNA (2 μg). Mean background (PCNA alone) subtracted counts are presented ± SD and comparisons made using student’s t-test. (C) Self methylation restricts activity. 2 μg of untreated C6orf211 or C6orf211 pretreated at 37°C for 90 min in the presence of 10 μM sinefungin were assayed for methyltransferase activity. Mean background (PCNA alone) subtracted absorbances from three replicate assays are presented. (D) PCNA is a target of C6orf211 methyltransferase activity. Mean background (PCNA alone) subtracted activities from three independent assays in the absence and presence of increasing amounts of PCNA are presented. (E) Pre-steady state kinetics support an inhibitory role for Armt1 self-methylation.