TY - JOUR
T1 - Identification of a cis-acting element of human dihydrofolate reductase mRNA
AU - Tai, Ningwen
AU - Schmitz, John C.
AU - Chen, Tian min
AU - O'Neill, Michelle B.
AU - Chu, Edward
N1 - Funding Information:
This work was supported, in part, by grants from the National Cancer Institute, USA (CA82897 and CA75712 to E.C.). The authors also thank the Developmental Therapeutics Program of the Yale Cancer Center and the VACT Cancer Center, VACT Healthcare System for their continued support of this research.
PY - 2008/5/9
Y1 - 2008/5/9
N2 - Human dihydrofolate reductase (DHFR) is a critical target in cancer chemotherapy. Previous studies showed that an 82-nt RNA fragment within the DHFR mRNA protein-coding region functions as a DHFR cis-acting response element. In this study, we further investigated the key elements contained within this sequence that are required for the DHFR mRNA-DHFR protein interaction. Using enzymatic foot-printing assays and RNA-binding experiments, we isolated a 27-nt sequence (DHFR27, corresponding to nts 407-433), which bound with high affinity and specificity to human DHFR to form a ribonucleoprotein complex. In vivo transient transfection experiments using a luciferase reporter system revealed that DHFR27 RNA could repress the luciferase expression in a DHFR-dependent manner when placed upstream of luciferase mRNA. This work provides new insights into the essential molecular elements that mediate RNA-protein interactions.
AB - Human dihydrofolate reductase (DHFR) is a critical target in cancer chemotherapy. Previous studies showed that an 82-nt RNA fragment within the DHFR mRNA protein-coding region functions as a DHFR cis-acting response element. In this study, we further investigated the key elements contained within this sequence that are required for the DHFR mRNA-DHFR protein interaction. Using enzymatic foot-printing assays and RNA-binding experiments, we isolated a 27-nt sequence (DHFR27, corresponding to nts 407-433), which bound with high affinity and specificity to human DHFR to form a ribonucleoprotein complex. In vivo transient transfection experiments using a luciferase reporter system revealed that DHFR27 RNA could repress the luciferase expression in a DHFR-dependent manner when placed upstream of luciferase mRNA. This work provides new insights into the essential molecular elements that mediate RNA-protein interactions.
KW - Dihydrofolate reductase
KW - RNA-protein interaction
KW - Translational regulation
KW - cis-acting element
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U2 - 10.1016/j.bbrc.2007.09.044
DO - 10.1016/j.bbrc.2007.09.044
M3 - Article
C2 - 18045573
AN - SCOPUS:41149124595
SN - 0006-291X
VL - 369
SP - 795
EP - 800
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -
