Proteome analysis reveals new mechanisms of Bcl11b-loss driven apoptosis.
The Bcl11b protein was shown to be important for a variety of functions such as T cell differentiation, normal development of central nervous system, and DNA damage response. Malignant T cells undergo apoptotic cell death upon BCL11B down-regulation, however, the detailed mechanism of cell death is not fully understood yet. Here we employed two-dimensional difference in-gel electrophoresis (2D-DIGE), mass spectrometry and cell biological experiments to investigate the role of Bcl11b in malignant T cell lines such as Jurkat and huT78. We provide evidence for the involvement of the mitochondrial apoptotic pathway and observed cleavage and fragments of known caspase targets such as myosin, spectrin, and vimentin. Our findings suggest an involvement of ERM proteins, which were up-regulated and phosphorylated upon Bcl11b down-regulation. Moreover, the levels of several proteins implicated in cell cycle entry, including DUT-N, CDK6, MCM4, MCM6, and MAT1 were elevated. Thus, the proteome data presented here confirm previous findings concerning the consequences of BCL11B knock-down and provide new insight into the mechanisms of cell death and cell cycle disturbances induced by Bcl11b depletion.
Interfakultäres Institut für Genetik und Funktionelle Genomforschung, Ernst-Moritz-Arndt-Universität Greifswald, Germany.
Narasimha Kumar
NK
Karanam
Piotr
P
Grabarczyk
Elke
E
Hammer
Christian
C
Scharf
Simone
S
Venz
Manuela
M
Gesell-Salazar
Winfried
W
Barthlen
Grzegorz K
GK
Przybylski
Christian A
CA
Schmidt
Uwe
U
Völker
D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D008928:Mitochondria / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D034741:RNA, Small Interfering / Q000235:genetics; D012097:Repressor Proteins / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism* / Q000473:pathology; D053719:Tandem Mass Spectrometry; D014157:Transcription Factors / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
C479740:BCL11B protein, human; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C086680:ETV5 protein, human; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins