The identification of thymidylate synthase peptide domains located in the interface region that bind thymidylate synthase mRNA.
Thymidylate synthase (TS) is a critical chemotherapeutic target and intracellular levels of TS are an important determinant of sensitivity to TS inhibitors. Translational autoregulation represents one cellular mechanism for controlling the level of expression of TS. This mechanism involves the binding of TS protein to its own messenger RNA (mRNA), thus, repressing translational efficiency. The presence of excess substrate or inhibitors of TS leads to derepression of protein binding to mRNA, resulting in increased translational efficiency and ultimately increased levels of TS protein. TS protein has been shown to bind to two distinct areas on its mRNA. The goal of the present work is to define the TS domains responsible for this interaction. Using a separate series of overlapping 17-mer peptides spanning the length of both the human and Escherichia coli TS sequences, we have identified six potential domains located in the interface region of the TS protein that bind TS mRNA. The identified domains that bind TS mRNA include three concordant regions in both the human and E. coli peptide series. Five of the six binding peptides contain at least one invariant arginine residue, which has been shown to be critical in other well-defined protein-RNA interactions. These data suggest that the identified highly conserved protein domains, which occur at the homodimeric interface of TS, represent potential participating sites for binding of TS protein to its mRNA.
Voeller
Donna M
DM
Center for Cancer Research, Basic Research Laboratory, National Cancer Institute, National Institutes of Health, Building 37 Room 6050D, 37 Convent Drive MSC 4255, Bethesda, MD 20892, USA. voellerd@mail.nih.gov
Zajac-Kaye
Maria
M
Fisher
Robert J
RJ
Allegra
Carmen J
CJ
D000595:Amino Acid Sequence; D001665:Binding Sites; D017124:Conserved Sequence; D004926:Escherichia coli / Q000201:enzymology*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D013940:Thymidylate Synthase / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D010446:Peptide Fragments; D012333:RNA, Messenger; D013940:Thymidylate Synthase