Abstract
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.
MeSH Terms
Adenosine Triphosphate/metabolism
Animals
Chromatography, Gel
Electrophoresis, Agar Gel/methods
Electrophoresis, Polyacrylamide Gel/methods
Heparin
Mammals
RNA Precursors/isolation & purification,metabolism
Ribonucleoproteins, Small Nuclear/isolation & purification
Spliceosomes/chemistry,metabolism
Chemicals
RNA Precursors
Ribonucleoproteins, Small Nuclear
Adenosine Triphosphate
Heparin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Das R
Harvard Medical School, Boston, Massachusetts 02115-5730, USA.
Reed R
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