Analysis of mRNA recognition by human thymidylate synthase

被引：18

作者：

Brunn, Nicholas D. ^{[1
]}

Dibrov, Sergey M. ^{[1
]}

Kao, Melody B. ^{[1
]}

Ghassemian, Majid ^{[1
]}

Hermann, Thomas ^{[1
,2
]}

机构：

[1] Univ Calif San Diego, Dept Chem & Biochem, La Jolla, CA 92093 USA

[2] Univ Calif San Diego, Ctr Drug Discovery Innovat, La Jolla, CA 92093 USA

来源：

BIOSCIENCE REPORTS | 2014年 / 34卷

基金：

美国国家卫生研究院;

关键词：

chemotherapy; translation regulation; UV cross-linking; X-ray crystallography; STRUCTURAL MECHANISM; REGULATORY ELEMENT; BINDING-SITE; AUTOREGULATION; 5-FLUOROURACIL; CONFORMATION; ENZYME; INHIBITION; REACTIVITY; RESISTANCE;

D O I：

10.1042/BSR20140137

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Expression of hTS (human thymidylate synthase), a key enzyme in thymidine biosynthesis, is regulated on the translational level through a feedback mechanism that is rarely found in eukaryotes. At low substrate concentrations, the ligand-free enzyme binds to its own mRNA and stabilizes a hairpin structure that sequesters the start codon. When in complex with dUMP (2'-deoxyuridine-5'-monophosphate) and a THF (tetrahydrofolate) cofactor, the enzyme adopts a conformation that is unable to bind and repress expression of mRNA. Here, we have used a combination of X-ray crystallography, RNA mutagenesis and site-specific cross-linking studies to investigate the molecular recognition of TS mRNA by the hTS enzyme. The interacting mRNA region was narrowed to the start codon and immediately flanking sequences. In the hTS enzyme, a helix-loop-helix domain on the protein surface was identified as the putative RNA-binding site.

引用

页码：905 / 913

页数：9

共 37 条

[1] The Phenix software for automated determination of macromolecular structures [J].

Adams, Paul D. ;

Afonine, Pavel V. ;

Bunkoczi, Gabor ;

Chen, Vincent B. ;

Echols, Nathaniel ;

Headd, Jeffrey J. ;

Hung, Li-Wei ;

Jain, Swati ;

Kapral, Gary J. ;

Kunstleve, Ralf W. Grosse ;

McCoy, Airlie J. ;

Moriarty, Nigel W. ;

Oeffner, Robert D. ;

Read, Randy J. ;

Richardson, David C. ;

Richardson, Jane S. ;

Terwilliger, Thomas C. ;

Zwart, Peter H. .

METHODS, 2011, 55 (01) :94-106

[2] The structural mechanism for half-the-sites reactivity in an enzyme, thymidylate synthase, involves a relay of changes between subunits [J].

Anderson, AC ;

O'Neil, RH ;

DeLano, WL ;

Stroud, RM .

BIOCHEMISTRY, 1999, 38 (42) :13829-13836

[3] THE CCP4 SUITE - PROGRAMS FOR PROTEIN CRYSTALLOGRAPHY [J].

BAILEY, S .

ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY, 1994, 50 :760-763

[4] Effects of ligand binding and conformational switching on intracellular stability of human thymidylate synthase [J].

Berger, SH ;

Berger, FG ;

Lebioda, L .

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 2004, 1696 (01) :15-22

[5] Targeting a Regulatory Element in Human Thymidylate Synthase mRNA [J].

Brunn, Nicholas D. ;

Sega, Emily Garcia ;

Kao, Melody B. ;

Hermann, Thomas .

CHEMBIOCHEM, 2012, 13 (18) :2738-2744

[6] THE CATALYTIC MECHANISM AND STRUCTURE OF THYMIDYLATE SYNTHASE [J].

CARRERAS, CW ;

SANTI, DV .

ANNUAL REVIEW OF BIOCHEMISTRY, 1995, 64 :721-762

[7] Thymidylate synthase inhibitors as anticancer agents: from bench to bedside [J].

Chu, E ;

Callender, MA ;

Farrell, MP ;

Schmitz, JC .

CANCER CHEMOTHERAPY AND PHARMACOLOGY, 2003, 52 (Suppl 1) :S80-S89

[8]

Chu E, 1996, Cancer Treat Res, V87, P175

[9] IDENTIFICATION OF AN RNA-BINDING SITE FOR HUMAN THYMIDYLATE SYNTHASE [J].

CHU, E ;

VOELLER, D ;

KOELLER, DM ;

DRAKE, JC ;

TAKIMOTO, CH ;

MALEY, GF ;

MALEY, F ;

ALLEGRA, CJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (02) :517-521

[10] AUTOREGULATION OF HUMAN THYMIDYLATE SYNTHASE MESSENGER-RNA TRANSLATION BY THYMIDYLATE SYNTHASE [J].

CHU, E ;

KOELLER, DM ;

CASEY, JL ;

DRAKE, JC ;

CHABNER, BA ;

ELWOOD, PC ;

ZINN, S ;

ALLEGRA, CJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (20) :8977-8981

← 1 2 3 4 →