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Biochemical analysis of TREX complex recruitment to intronless and intron‐containing yeast genes
Author(s) -
Abruzzi Katharine Compton,
Lacadie Scott,
Rosbash Michael
Publication year - 2004
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1038/sj.emboj.7600261
Subject(s) - biology , intron , gene , genetics , yeast , computational biology
The TREX complex is involved in both transcription elongation and mRNA export and is recruited to nascent transcription complexes. We have examined Yra1p, Sub2p and Hpr1p recruitment to nine genes of varying lengths and transcription frequencies. All three proteins increase from the 5′ to the 3′ ends of the four intronless genes examined. A modified chromatin immunoprecipitation assay that includes an RNase step indicates that Sub2p is bound to nascent RNA, Yra1p is associated with both RNA and DNA, and Hpr1p is associated with DNA. Although Hpr1p is recruited similarly to both intronless and intron‐containing genes, low Yra1p and Sub2p levels are present on a subset of intron‐containing genes. The residual Yra1p and Sub2p recruitment is less RNA‐associated, and this correlates with high levels of U1 SnRNP on these genes. These experiments support a model in which TREX is recruited via the transcription machinery and then Yra1p and Sub2p are transferred to the nascent RNA. On some intron‐containing genes, retention and/or transfer of Yra1p and Sub2p to nascent RNA are inhibited.