Low temperature enhancement of reporter genes expression directed by human immunodeficiency virus type 1 long terminal repeat.
Bacteria and eukaryotic cells respond to cold stress by inducing and enhancing the synthesis of specific arrays of proteins. We describe here cold-induced enhancement of expression for two reporter genes; luciferase and beta-galactosidase, both under the control of HIV-1 LTR sequences, observed in mouse fibroblasts and human HeLa cells respectively. Increased expression of luciferase in fibroblasts when shifted to 25 degrees C was detectable at 30 degrees C but was not observed following cold shock at 4 degrees C. To sustain the cold-induced effect, cells had to be kept at subphysiological temperature. The observed enhancement of luciferase activity did not result from a particular site of integration of the reporter gene and was evident whether cold-stressed cells were stationary or growing. Cold-induced expression of luciferase was evidenced at the protein level, enzymatic activity and RNA level, furthermore, active transcription and translation were required for overexpression. The cold effect which has been generalized with the reporter gene beta-galactosidase appears to be a process involving, at least in part, the HIV-1 LTR sequences and might correspond to an increase in the half-life of mRNA. The cold-dependent enhanced expression of luciferase and beta-galactosidase reported here, together with data describing the activation of HIV-1 LTR by hyperthermia, point out the particular temperature sensitivity of these regulatory sequences. This potential thermal modulation may be useful in the comprehension of regulatory processes in latency and reactivation of viral expression during HIV-1 infection.
Chevrier-Miller
M
M
Ecole Normale Supérieure, CNRS-URA 1302, Paris, France.
Morange
M
M
Arrigo
A P
AP
Pinto
M
M
D016475:3T3 Cells; D000818:Animals; D003080:Cold Temperature; D004791:Enzyme Inhibitors; D015967:Gene Expression Regulation, Viral; D017930:Genes, Reporter; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics; D018922:HL-60 Cells; D006801:Humans; D008156:Luciferases / Q000037:antagonists & inhibitors / Q000235:genetics*; D051379:Mice; D012333:RNA, Messenger / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D008156:Luciferases; D001616:beta-Galactosidase