[RNA interference in Escherichia coli cells: the expression of molecules that are complementary to the lon gene mRNA in parallel orientation].
To study the effect of RNA interference (RNAi) on the activity of gene lon in Escherichia coli, genetic constructs were used that could express RNA molecules complementary to the 5' region of lon mRNA in the same direction. These RNAs were termed parallel RNAs (pRNAs). Two approaches were used to control expression. In one approach, lon gene activity was estimated genetically, based on the effect of the Lon protease on bioluminescence determined by the Vibrio fischeri lux regulon. The other approach was direct testing of ATP-dependent proteolysis in vitro. It was found that pRNA considerably suppressed lon expression. The antiparallel RNA (apRNA) was a less effective suppressor of this gene. The specific RNAi was found to decay gradually by the 40th generation. The data obtained indicate that Eubacterium cells have mechanisms for specific regulation of gene activity that are sensitive to the formation of both parallel and antiparallel RNA duplexes involving mRNA of the given gene.
Churikov
N A
NA
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia. tchur@imb.ac.ru
Chistiakova
L G
LG
Zavil'gel'skiĭ
G B
GB
Manukhov
I V
IV
D049069:ATP-Dependent Proteases; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics*; D049070:Protease La; D012333:RNA, Messenger / Q000235:genetics*; D012697:Serine Endopeptidases / Q000235:genetics*
D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases; C483567:Lon protein, E coli; D049070:Protease La