Resolution of the mammalian E complex and the ATP-dependent spliceosomal complexes on native agarose mini-gels.
A great deal of progress in elucidating the mechanisms of spliceosome assembly has been achieved by analyzing the A, B, and C spliceosomal complexes on native polyacrylamide gels. In contrast, progress in understanding the earliest spliceosomal complex E has been hampered because this complex dissociates on native gels and is difficult to detect by other methods. Here we report conditions for resolving the spliceosomal complex E using a native horizontal agarose mini-gel system. This system also provides a simple alternative to polyacrylamide gels for resolving the ATP-dependent spliceosomal complexes.
Das
R
R
Harvard Medical School, Boston, Massachusetts 02115-5730, USA.
Reed
R
R
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002850:Chromatography, Gel; D004587:Electrophoresis, Agar Gel / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006493:Heparin; D008322:Mammals; D012322:RNA Precursors / Q000302:isolation & purification / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000302:isolation & purification*; D017381:Spliceosomes / Q000737:chemistry* / Q000378:metabolism*
D012322:RNA Precursors; D017411:Ribonucleoproteins, Small Nuclear; D000255:Adenosine Triphosphate; D006493:Heparin