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. 2010 Dec 13;286(6):4931–4940. doi: 10.1074/jbc.M110.195024

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FIGURE 6. — Effect of Nej1 on Dnl4 deadenylation and readenylation. A, His-tagged Dnl4-Lif1 (1 pmol) was preincubated without (lanes 1 and 2) or with (2 pmol; lanes 3 and 4) Nej1. B, His-tagged Dnl4-Lif1 (1 pmol) was preincubated without (lanes 1 and 2) or with (2 pmol; lanes 3 and 4) Nej1 in the presence of 10 mm pyrophosphate. C, His-tagged Dnl4-Lif1 (1 pmol) was preincubated without (lanes 1 and 2) or with (2 pmol; lanes 3 and 4) Nej1 in the presence of linear DNA oligonucleotide duplex with cohesive ligatable ends (100 pmol). After collection on nickel beads, Dnl4-Lif1 was incubated with [α-³²P]ATP in the absence (lanes 1 and 4) or presence (2 pmol; lanes 2 and 3). of Nej1 Proteins were separated by SDS-PAGE and labeled. Dnl4 was detected and quantitated by PhosphorImager analysis. The results of three independent experiments are shown graphically. Error bars indicate mean ± S.D.