Open AccessCorrect removal by splicing of aNeurosporaintron in yeast
Author(s) -
Lambertus P. Woudt,
Johannes Jacobus van den Heuvel,
Mary M.C. van Raamsdonk-Duin,
Willem H. Mager,
Rudi J. Planta
Publication year - 1985
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/13.21.7729
Subject(s) - biology , intron , neurospora , rna splicing , genetics , yeast , group i catalytic intron , group ii intron , computational biology , gene , neurospora crassa , rna , mutant
Processing of intron-containing nuclear messenger RNAs in yeast require an internal conserved sequence (ICS) element, UACUAAC. Similar elements (ugCUAGAC) have been identified in sequences interrupting nuclear genes of the related ascomycete Neurospora crassa. To examine the structural splicing requirements in yeast, we constructed hybrid genes containing the intron of the Neurospora histone H3 gene and cloned them into high copy number yeast vectors. Subsequently we analyzed the RNAs transcribed in yeast from the fusion genes by Northern analysis and primer extended sequencing. It turned out that the Neurospora intron, which contains the sequence element UGCUAAC, can be removed, though very inefficiently, provided that it is located near the 5'-end of the primary transcript. This proves that an A at the second position of the ICS is no absolute requirement for splicing in yeast. In addition, the results indicate that the yeast splicing machinery is intron-position dependent.