MOLECULAR BASIS FOR TS TRANSLATIONAL REGULATION

Project: Research project

Project Details

Description

DESCRIPTION: (Applicant's Abstract) The long-term goal of this project is to
investigate the translational regulation of cellular gene expression. As his
model system, the applicant will elucidate the translational regulation of
thymidylate synthase (TS) expression, as TS is a critical target in cancer
chemotherapy. This enzyme catalyzes the folate-dependent reductive methylation
reaction which provides for the sole intracellular de novo source of
thymidylate, a key precursor for DNA biosynthesis, and thus, TS plays a central
role in maintaining the metabolic requirements of the cell. Studies from the
applicant's laboratory have shown that TS also functions as an RNA binding
protein in which it binds with high affinity (1-3 nM) to two different sites on
its own TS mRNA. One site is located in the 5'-untranslated region and the
second site in the protein-coding region. Binding of TS to either element
results in translational repression of TS mRNA and inhibition of synthesis of
TS protein. The model of TS translational autoregulation appears to have
biological relevance in that it offers a rational mechanism for the tight
control of TS expression within a given cell. However, treatment of TS protein
with inhibitor compounds such as FdUMP or the antifolate inhibitor D1694 alters
the synthesis of new TS protein. Disruption of this regulatory process would
appear to provide an efficient adaptive mechanism for malignant cells to
protect themselves in response to exposure to cytotoxic stress and thereby
result in the development of cellular resistance. To further our understanding
of the molecular elements underlying the translational regulation of TS, three
specific aims are proposed in this application: (1) Characterize in further
detail the critical cis-acting elements on TS mRNA that are required for the TS
mRNA-TS protein for protein recognition of both the 5'-upstream and
protein-coding region binding sites; (2) Characterize in further detail the
critical domain or domains on the TS protein as well as identify the specific
amino acid contact points directly involved in RNA binding; and (3)
Characterize the intracellular localization of the TS protein-TS mRNA
ribonucleoprotein complex. These studies will provide enhanced insight into the
molecular elements mediating the interaction between TS protein and its cognate
TS mRNA. Moreover, these molecular-based studies may provide the rational basis
for the development of novel therapeutic strategies for human cancer.
StatusFinished
Effective start/end date9/18/971/31/07

Funding

  • National Cancer Institute: $245,050.00
  • National Cancer Institute
  • National Cancer Institute: $245,050.00

ASJC

  • Genetics
  • Molecular Biology
  • Cancer Research

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