Fig. 2.

ATRIP 108–791 associates directly with RPA–ssDNA. (A) Full-length ATRIP (ATRIP 1–791), ATRIP 108–791, and ATRIP 198–791 were synthesized by using coupled in vitro transcription and translation in rabbit reticulocyte lysate and labeled with [³⁵S]methionine. The reticulocyte lysates containing the in vitro-translated proteins were incubated with ssDNA or RPA–ssDNA. The in vitro-translated proteins present in the input (5%) or associated with ssDNA or RPA–ssDNA were detected by autoradiograph. (B) The binding of the in vitro-translated full-length ATRIP and ATRIP 108–791 to RPA–ssDNA was analyzed in the same buffer as described above (buffer A) and in NETN buffer (buffer B). The proteins that were present in the input (10%) or associated with RPA–ssDNA were detected by autoradiography. (C) Purified GST–ATRIP 108–791 on a Coomassie blue-stained SDS/PAGE. (D) We incubated ≈0.5 μgof purified GST–ATRIP 108–791 and 10 μg of purified GST with ssDNA or RPA–ssDNA, respectively. The proteins that were present in the input (10%) or associated with ssDNA or RPA–ssDNA were detected by using anti-GST antibody.