Abstract:
:The pre-mRNA 5' splice site is recognized by the ACAGA box of U6 spliceosomal RNA prior to catalysis of splicing. We previously identified a mutant U4 spliceosomal RNA, U4-cs1, that masks the ACAGA box in the U4/U6 complex, thus conferring a cold-sensitive splicing phenotype in vivo. Here, we show that U4-cs1 blocks in vitro splicing in a temperature-dependent, reversible manner. Analysis of splicing complexes that accumulate at low temperature shows that U4-cs1 prevents U4/U6 unwinding, an essential step in spliceosome activation. A novel mutation in the evolutionarily conserved U5 snRNP protein Prp8 suppresses the U4-cs1 growth defect. We propose that wild-type Prp8 triggers unwinding of U4 and U6 RNAs only after structurally correct recognition of the 5' splice site by the U6 ACAGA box and that the mutation (prp8-201) relaxes control of unwinding.
journal_name
Mol Celljournal_title
Molecular cellauthors
Kuhn AN,Li Z,Brow DAdoi
10.1016/s1097-2765(00)80175-6subject
Has Abstractpub_date
1999-01-01 00:00:00pages
65-75issue
1eissn
1097-2765issn
1097-4164pii
S1097-2765(00)80175-6journal_volume
3pub_type
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