title: The C-terminal transmembrane domain of Bcl-xL mediates changes in mitochondrial morphology.
abstract: We investigate the effect of mitochondrial localization and the Bcl-x(L) C-terminal transmembrane (TM) domain on mitochondrial morphology and subcellular light scattering. CSM 14.1 cell lines stably expressed yellow fluorescent protein (YFP), YFP-Bcl-x(L,) YFP-Bcl-x(L)-DeltaTM, containing the remainder of Bcl-x(L) after deletion of the last 21 amino acids corresponding to the TM domain, or YFP-TM, consisting of YFP fused at its C-terminal to the last 21 amino acids of Bcl-x(L). YFP-Bcl-x(L) and YFP-TM localized to the mitochondria. Their expression decreased the intensity ratio of wide-to-narrow angle forward scatter by subcellular organelles, and correlated with an increase in the proportion of mitochondria with an expanded matrix having greatly reduced intracristal spaces as observed by electron microscopy. Cells expressing YFP-TM also exhibited significant autophagy. In contrast, YFP-Bcl-x(L)-DeltaTM was diffusely distributed in the cells, and its expression did not alter light scattering or mitochondrial morphology compared with parental cells. Expression of YFP-Bcl-x(L) or YFP-Bcl-x(L)-DeltaTM provided significant resistance to staurosporine-induced apoptosis. Surprisingly however, YFP-TM expression also conferred a moderate level of cell death resistance in response to staurosporine. Taken together, our results suggest the existence of a secondary Bcl-x(L) function that is mediated by the transmembrane domain, alters mitochondrial morphology, and is distinct from BH3 domain sequestration.
authors.affiliation: Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA.
authors.forename: Jing-Yi
authors.initials: JY
authors.lastname: Zheng
authors.forename: Yien-Che
authors.initials: YC
authors.lastname: Tsai
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kadimcherla
authors.forename: Rong
authors.initials: R
authors.lastname: Zhang
authors.forename: Julia
authors.initials: J
authors.lastname: Shi
authors.forename: George A
authors.initials: GA
authors.lastname: Oyler
authors.forename: Nada N
authors.initials: NN
authors.lastname: Boustany
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D048429:Cell Size; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure*; D009474:Neurons / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D017434:Protein Structure, Tertiary; D051381:Rats; D051020:bcl-X Protein / Q000378:metabolism*
chemical_list: C494813:Bcl2l1 protein, rat; D051020:bcl-X Protein