title: New methods to investigate ATP requirement for pre-mRNA splicing: inhibition by hexokinase/glucose or an ATP-binding site blocker.
abstract: Addition of yeast hexokinase and glucose at various time points of the pre-mRNA splicing reaction rapidly depleted ATP and inhibited further progress of the reaction, indicating that ATP is required for both the first and second steps of splicing. ATP analogues, p-fluorosulfonylbenzoyl-5'-adenosine (FSBA) and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl), which can modify amino acids at the ATP-binding site of a protein, inactivated the splicing activity of the nuclear extract. While the inactivation by the former was irreversible, the splicing activity was complemented by a Micrococcal nuclease-treated extract. This ATP analogue (FSBA) may be a useful tool for identification of ATP-dependent splicing factors.
authors.affiliation: Department of Biology, Faculty of Science, Kyushu University, Fukuoka.
authors.forename: K
authors.initials: K
authors.lastname: Tatei
authors.forename: K
authors.initials: K
authors.lastname: Kimura
authors.forename: Y
authors.initials: Y
authors.lastname: Ohshima
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D006593:Hexokinase / Q000494:pharmacology; D008722:Methods; D058914:Purinergic Antagonists; D012326:RNA Splicing; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D058914:Purinergic Antagonists; D012333:RNA, Messenger; D000255:Adenosine Triphosphate; D006593:Hexokinase; D005947:Glucose