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According to our matching algorithm, William Mattox is the likely recipient of the following grants.
Years |
Recipients |
Code |
Title / Keywords |
Matching score |
1994 — 1998 |
Mattox, William W |
R29Activity Code Description: Undocumented code - click on the grant title for more information. |
Regulating Alternative Pre-Mrna Splicing in Drosophila @ University of Texas Md Anderson Can Ctr
Alternative pre-mRNA splicing is used widely among eucaryotes in the control of both gene expression and function. Regulation of splicing plays a critical role in a wide variety of processes needed for normal development. Despite its importance in gene regulation, remarkably little is known about the molecular mechanisms that result in alternative splicing. Particularly lacking is information on the biochemical activities of the trans-acting regulators that are thought to direct usage of alternative splicing pathways. Very few such regulators have yet been identified and, of those known, only a few have been studied in sufficient detail to yield significant information about their mechanisms of action. We propose to investigate the molecular mechanism by which transformer-2 (tra-2), an established splicing regulator from Drosophila melanogaster, functions to repress the splicing of an intron found within the tra-2 pre- mRNA itself. We will first investigate the physical interaction of tra-2 protein and pre-mRNA by using a UV crosslinking assay to identify both RNA and protein sequences needed for binding. The significance of the identified binding sites with respect to regulated splicing will be evaluated in the developmental context of the fly's male germline where autoregulation normally occurs. This will be accomplished in experiments using stable transgenic fly strains in which the binding site sequences have been altered. Additional studies will follow up on our preliminary finding that indicate that a general factor plays an important role in this binding interaction. We describe an approach for both identifying this factor and assessing its role in regulation of splicing. Finally, we will use a cell free in vitro splicing system to examine how tra-2 protein molecules interfere with the splicing of the M1 intron. The aim of these studies will be to identify the specific interactions between the splicing machinery and the pre-mRNA that are blocked in the presence of RNA bound tra-2 protein molecules. The results of these studies will provide substantial insights into the mechanism by which tra-2 protein molecules affect splicing. Such insights should prove helpful in understanding how splicing regulators function to control alternative splicing in other systems.
|
0.907 |
1999 — 2002 |
Mattox, William W |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Phosphorylation and the Regulation of Rna Splicing @ University of Texas Md Anderson Can Ctr
Alternative pre-mRNA splicing affects the expression of gene products involved in many aspects of cell growth and differentiation, yet the mechanisms by which it is regulated are only beginning to be elucidated. SR proteins and related factors are thought to play important roles in the regulation of splicing. These factors form regulatory complexes at sites internal to the exons of alternatively spliced pre-mRNAs and function to recruit factors from the general splicing machinery to nearby splice sites that otherwise would not be recognized. Although SR proteins are highly phosphorylated, the role of phosphorylation in the regulation of splicing is largely unknown. We propose to investigate how SR protein phosphorylation affects the assembly and function of sex-specific regulatory complexes formed on a splicing enhancer in the doublesex pre-mRNA. Preliminary studies indicate that the LAMMER kinase Darkener-of- apricot phosphorylates SR proteins and is required for normal regulation of doublesex RNA splicing. We will test whether the phosphorylation of proteins from the doublesex splicing enhancer complex affects the ability of SR proteins, and related regulatory factors such as TRA and TRA2, to interact with each other and with the pre-mRNA. Other experiments will compare the effects of different classes of kinases on SR protein function. Finally, we will produce strains of Drosophila with mutations in SR protein kinases to examine their roles in development and on the phosphorylation of specific SR proteins in vivo. We anticipate that these studies will provide significant new insights into the mechanisms by which splicing is regulated and will open up new avenues for future studies on phosphorylation of SR proteins.
|
0.907 |