The impact of tyrosine kinase 2 (Tyk2) on the proteome of murine macrophages and their response to lipopolysaccharide (LPS).
Tyrosine kinase 2 (Tyk2) belongs to the Janus kinase (Jak) family and is involved in signalling via a number of cytokines. Tyk2-deficient mice are highly resistant to lipopolysaccharide (LPS)-induced endotoxin shock. Macrophages are key players in the pathogenesis of endotoxin shock and, accordingly, defects in the LPS responses of Tyk2(-/-) macrophages have been reported. In the present study, the molecular role of Tyk2 is investigated in more detail using a proteomics approach. 2-D DIGE was applied to compare protein patterns from wild-type and Tyk2(-/-) macrophages and revealed significant differences in protein expression patterns between the genotypes before and after LPS treatment. Twenty-one proteins deriving from 25 differentially expressed spots were identified by MALDI/ESI MS. Among them, we show for N-myc interactor that its mRNA transcription/stability is positively influenced by Tyk2. In contrast, LPS-induced expression of plasminogen activator 2 protein but not mRNA is strongly enhanced in the absence of Tyk2. Our data furthermore suggest an influence of Tyk2 on the subcellular distribution of elongation factor 2 and on LPS-mediated changes in the peroxiredoxin 1 spot pattern. Thus, our results imply regulatory roles of Tyk2 at multiple levels and establish novel connections between Tyk2 and several cellular proteins.
Radwan
Marta
M
Institute of Animal Breeding and Genetics, University of Veterinary Medicine Vienna, Vienna, Austria.
Miller
Ingrid
I
Grunert
Tom
T
Marchetti-Deschmann
Martina
M
Vogl
Claus
C
O'Donoghue
Niaobh
N
Dunn
Michael J
MJ
Kolbe
Thomas
T
Allmaier
Günter
G
Gemeiner
Manfred
M
Müller
Mathias
M
Strobl
Birgit
B
D000818:Animals; D018797:Cell Cycle Proteins / Q000096:biosynthesis; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D020652:Peptide Elongation Factor 2 / Q000096:biosynthesis; D054464:Peroxiredoxins / Q000096:biosynthesis; D017396:Plasminogen Activator Inhibitor 2 / Q000096:biosynthesis; D020543:Proteome / Q000187:drug effects*; D012333:RNA, Messenger / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053634:TYK2 Kinase / Q000172:deficiency / Q000502:physiology*; D000099259:N-myc Downstream-Regulated Gene 1 Protein
D018797:Cell Cycle Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D020652:Peptide Elongation Factor 2; D054464:Peroxiredoxins; D017396:Plasminogen Activator Inhibitor 2; D020543:Proteome; D012333:RNA, Messenger; D053634:TYK2 Kinase; D000099259:N-myc Downstream-Regulated Gene 1 Protein; C489899:Prdx1 protein, mouse; C499356:Tyk2 protein, mouse