PMID- 10201640
OWN - NLM
STAT- MEDLINE
DCOM- 19990421
LR  - 20190701
IS  - 0024-3205 (Print)
IS  - 0024-3205 (Linking)
VI  - 64
IP  - 11
DP  - 1999
TI  - Mu-opioid receptor expression in High Five insect cells is regulated by 5' 
      untranslated region (5'UTR).
PG  - 913-21
AB  - There is increasing evidence that the 5'UTR of mRNAs affects regulation of gene 
      expression in eukaryotic cells. We examined the overexpression of the mu-opioid 
      receptor in High Five insect cells, employing rat mu-receptor cDNA linked to 
      variable lenghts of their native 5'UTR. The sequences employed consist of either 
      209 nucleotides (termed ,,long") upstream the translation initiation site of the 
      mu-receptor mRNA, or a truncated 5'UTR comprising only 11 nucleotides (,,short"). 
      These constructs served to generate recombinant baculovirus for the expression of 
      mu-receptor protein in High Five insect cells. 48 hours after baculovirus 
      infection cells were harvested for mu-receptor characterization or RNA analysis. 
      Scatchard analysis of radioligand binding consistently revealed three to four 
      fold higher concentrations of the mu-opioid receptors expressed with the ,,long" 
      over the ,,short" UTR containing baculovirus. The distinct expression rates of 
      mu-receptors paralleled the amounts of mRNAs determined by RNase protection 
      assay. Regardless of the distinct 5'UTR regions, the expressed opioid receptors 
      displayed identical high affinity binding characteristics for the opioid 
      antagonist diprenorphine and similar EC50 values to inhibit forskolin (10(-5) M) 
      stimulated cAMP synthesis. Our results demonstrate that the native 5'UTR of the 
      mu-opioid receptor has an enhancing effect on expression in the 
      baculovirus/insect cell system.
FAU - Belke-Louis, G F
AU  - Belke-Louis GF
AD  - Institute of Pharmacology, Toxicology and Pharmacy, Ludwig Maximilians 
      University, Muenchen, Germany. Belke_Louis@pharmtox.vetmed.uni-muenchen.de
FAU - Wehmeyer, A
AU  - Wehmeyer A
FAU - Schulz, R
AU  - Schulz R
LA  - eng
PT  - Journal Article
PL  - Netherlands
TA  - Life Sci
JT  - Life sciences
JID - 0375521
RN  - 0 (5' Untranslated Regions)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Opioid, mu)
RN  - E0399OZS9N (Cyclic AMP)
SB  - IM
MH  - 5' Untranslated Regions/*physiology
MH  - Animals
MH  - Baculoviridae/genetics
MH  - Cyclic AMP/biosynthesis
MH  - Insecta
MH  - RNA, Messenger/analysis
MH  - Rats
MH  - Receptors, Opioid, mu/biosynthesis/*genetics
EDAT- 1999/04/14 00:00
MHDA- 1999/04/14 00:01
CRDT- 1999/04/14 00:00
PHST- 1999/04/14 00:00 [pubmed]
PHST- 1999/04/14 00:01 [medline]
PHST- 1999/04/14 00:00 [entrez]
AID - S002432059900017X [pii]
AID - 10.1016/s0024-3205(99)00017-x [doi]
PST - ppublish
SO  - Life Sci. 1999;64(11):913-21. doi: 10.1016/s0024-3205(99)00017-x.