Structure of translation initiation region of c-myc mRNA and thermostability of the c-myc mRNA/antisense ODN complex.
We enzymatically synthesized c-myc mRNA fragment containing the translation initiation (TI) region (36mer) and examined the structure by CD, 1H-NMR and RNaseV1 digestion. These study demonstrated that the c-myc mRNA fragment forms a hairpin structure at 37 degrees C. Six 15-mer antisense oligodeoxynucleotides(ODNs), which hybridize to the c-myc mRNA fragment were designed, and synhesized. Thermostabilities of the antisense ODNs-mRNA fragment complexes were examined. Antisense A (complementary to the c-myc mRNA from -9 to +6)-c-myc mRNA fragment complex had the highest melting temperature(Tm), and Antisense D (complementary to the c-myc mRNA from +1 to +15)-c-myc mRNA fragment complex had the most different Tm from a predicted Tm. Furthermore, Antisense D exhibited high inhibitory effect on HL60 cell proliferation, whereas Antisense A did not. The results suggest that the difference in the antisense effect is not simply due to the complex thermostability.
Tanaka
T
T
Department of Bioengineering, Faculty of Engineering, Yokohama National University, Japan.
Kurihara
Y
Y
Sakamoto
T
T
Furusawa
M
M
Kobayashi
A
A
Katahira
M
M
Takeuchi
K
K
Sugimoto
N
N
Uesugi
S
S
D002460:Cell Line; D004355:Drug Stability; D016259:Genes, myc; D006801:Humans; D015394:Molecular Structure; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D016376:Oligonucleotides, Antisense / Q000737:chemistry* / Q000235:genetics*; D010442:Peptide Chain Initiation, Translational / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry* / Q000235:genetics*; D013696:Temperature
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger