Loss of binding of cytoplasmic SAMHD1 K11A to SAMHD1-associated nuclear proteins.

<p>(A) HEK293T cells were transfected with SAMHD1 WT, K11A, Δ2–109, or empty vector control, respectively. Co- immunoprecipitation was carried out 48 h later by using anti-HA affinity matrix (Roche). Protein samples were analyzed by mass-spec as described in the Methods. (B) Interaction between SAMHD1 WT/mutant proteins and KPNB1. HEK293T cells were transfected with a control vector (lane 1, VR) or WT SAMHD1-HA (lane 2), or one of the indicated SAMHD1-HA mutants (lanes 3–4). Cell lysates were prepared 48 h after transfection and immunoprecipitated with anti-HA affinity matrix (Roche). The interaction of KPNB1 with WT or mutated SAMHD1-HA molecules was detected by immunoblotting with anti-HA antibody to detect SAMHD1-HA. A specific anti-KPNB1 was used to detect endogenous KPNB1.