Lindsey-Boltz, Laura A and Chawla, Geetanjali and Srinivasan, N and Vijeyraghavan, Usha and Mariano, A and Garica-Blanco, * (2000) The carboxy terminal WD domain of the pre-mRNA splicing factor Prp17p is critical for function. In: RNA, 6 (9). pp. 1289-1305.
The_carboxy.pdf - Published Version
Restricted to Registered users only
Download (4Mb) | Request a copy
In Saccharomyces cerevisiae, Prp17p is required for the efficient completion of the second step of pre-mRNA splicing. The function and interacting factors for this protein have not been elucidated. We have performed a mutational analysis of yPrp17p to identify protein domains critical for function. A series of deletions were made throughout the region spanning the N-terminal 158 amino acids of the protein, which do not contain any identified structural motifs. The C-terminal portion (amino acids 160–455) contains a WD domain containing seven WD repeats. We determined that a minimal functional Prp17p consists of the WD domain and 40 amino acids N-terminal to it. We generated a three-dimensional model of the WD repeats in Prp17p based on the crystal structure of the [beta]-transducin WD domain. This model was used to identify potentially important amino acids for in vivo functional characterization. Through analysis of mutations in four different loops of Prp17p that lie between [beta] strands in the WD repeats, we have identified four amino acids, 235TETG238, that are critical for function. These amino acids are predicted to be surface exposed and may be involved in interactions that are important for splicing. Temperature-sensitive prp17 alleles with mutations of these four amino acids are defective for the second step of splicing and are synthetically lethal with a U5 snRNA loop I mutation, which is also required for the second step of splicing. These data reinforce the functional significance of this region within the WD domain of Prp17p in the second step of splicing.
|Item Type:||Journal Article|
|Additional Information:||Copyright of this article belongs to Cambridge University Press.|
|Keywords:||b propeller;homology modeling;mutational analysis;nuclear localization;Prp17;second-step splicing; U5 snRNA|
|Department/Centre:||Division of Biological Sciences > Microbiology & Cell Biology|
|Date Deposited:||15 Sep 2011 04:29|
|Last Modified:||15 Sep 2011 04:29|
Actions (login required)