Gene expression analysis of murine cells producing amphotropic mouse leukaemia virus at a cultivation temperature of 32 and 37 degrees C.
Cultivation of retrovirus packaging cells at 32 degrees C represents a common procedure to achieve high titres in mouse retrovirus production. Gene expression profiling of mouse NIH 3T3 cells producing amphotropic mouse leukaemia virus 4070A revealed that 10 % of the 1176 cellular genes investigated were regulated by temperature shift (37/32 degrees C), while 5 % were affected by retrovirus infection. Strikingly, retrovirus production at 32 degrees C activated the cholesterol biosynthesis/transport pathway and caused an increase in plasma membrane cholesterol levels. Furthermore, these conditions resulted in transcriptional activation of smoothened (smo), patched (ptc) and gli-1; Smo, Ptc and Gli-1, as well as cholesterol, are components of the Sonic hedgehog (Shh) signalling pathway, which directs pattern formation, diversification and tumourigenesis in mammalian cells. These findings suggest a link between cultivation at 32 degrees C, production of MLV-A and the Shh signalling pathway.
Beer
Christiane
C
Molecular Biotechnology, German Research Centre for Biotechnology, GBF, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
Buhr
Petra
P
Molecular Biotechnology, German Research Centre for Biotechnology, GBF, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
Hahn
Heidi
H
Human Genetics, Georg August University, Göttingen, Germany.
Laubner
Daniela
D
Institute for Experimental Genetics, GSF-National Research Centre for Environment and Health, Neuherberg, Germany.
Wirth
Manfred
M
Molecular Biotechnology, German Research Centre for Biotechnology, GBF, Mascheroder Weg 1, D-38124 Braunschweig, Germany.
D016475:3T3 Cells / Q000502:physiology / Q000821:virology*; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002784:Cholesterol / Q000032:analysis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D053823:Hedgehog Proteins; D009052:Leukemia Virus, Murine / Q000472:pathogenicity / Q000502:physiology*; D051379:Mice; D020411:Oligonucleotide Array Sequence Analysis; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013696:Temperature; D015534:Trans-Activators / Q000378:metabolism; D019065:Virus Assembly
D053823:Hedgehog Proteins; D011506:Proteins; C465570:Shh protein, mouse; D015534:Trans-Activators; D002784:Cholesterol