Transcriptomics

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Structural evidence for consecutive Hel308-like modules in the spliceosomal ATPase Brr2


ABSTRACT: Brr2 is a DExD/H-box helicase responsible for U4/U6 unwinding, a critical step in spliceosomal activation. Brr2 contains an N-terminal domain and two tandem sets of a helicase-like domain followed by a Sec63 domain with unknown function. We determined the crystal structure of the second Sec63 domain, which unexpectedly resembles domains 4 and 5 of DNA helicase Hel308. The helicase-like domain upstream of Sec63 has clear sequence similarity with domains 1-3 of Hel308. In addition modeling indicates that Brr2 is composed of an N-terminal domain and two consecutive Hel308-like modules (Hel308-I and II). Together this provides our first glimpse of the overall structure of this large and unique spliceosomal ATPase and helicase. Our structural model and mutagenesis data suggest that Brr2 shares a similar helicase mechanism to Hel308, that differs from many DEAD-box proteins. We demonstrate that Hel308-II interacts with Prp8 and Snu114 in vitro and in vivo, potentially serving as a mediator for the regulation of Brr2’s activity by Prp8. We further find that the C-terminal region of Prp8 (Prp8-CTR) facilitates the binding of the Brr2/Prp8-CTR complex to U4/U6, suggesting a potential role of Prp8-CTR as an auxiliary substrate binding and specificity domain for Brr2. Splicing specific microarrays were used to assess the genome-wide defects in pre-mRNA splicing that result from a deletion of the second Sec63 domain of yeast Brr2.

ORGANISM(S): Saccharomyces cerevisiae  

SUBMITTER: Laura-Oana Bud  Lingdi Zhang  Jay Nix   Jeffrey A Pleiss   Rui Zhao   James Shanks   Christine Guthrie   Tao Xu   Corina Maeder      

PROVIDER: E-GEOD-16135 | ArrayExpress | 2010-05-18

SECONDARY ACCESSION(S): GSE16135PRJNA115033

REPOSITORIES: GEO, ArrayExpress

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Structural evidence for consecutive Hel308-like modules in the spliceosomal ATPase Brr2.

Zhang Lingdi L   Xu Tao T   Maeder Corina C   Bud Laura-Oana LO   Shanks James J   Nix Jay J   Guthrie Christine C   Pleiss Jeffrey A JA   Zhao Rui R  

Nature structural & molecular biology 20090614 7


Brr2 is a DExD/H-box helicase responsible for U4/U6 unwinding during spliceosomal activation. Brr2 contains two helicase-like domains, each of which is followed by a Sec63 domain with unknown function. We determined the crystal structure of the second Sec63 domain, which unexpectedly resembles domains 4 and 5 of DNA helicase Hel308. This, together with sequence similarities between Brr2's helicase-like domains and domains 1-3 of Hel308, led us to hypothesize that Brr2 contains two consecutive He  ...[more]

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