Transforming growth factor beta (TGF beta)-induced nuclear localization of apolipoprotein J/clusterin in epithelial cells.
Apolipoprotein J (apoJ)/clusterin was first identified as an 80 kDa secretory glycoprotein present in most body fluids. It has been implicated in a variety of physiological processes including cellular differentiation and apoptosis. We demonstrate here that in addition to the well characterized secreted form of the protein, there exists an intracellular, nuclear form of apoJ. This intracellular form of the protein is induced to accumulate in the nucleus of two epithelial cell lines (HepG2 and CCL64) in response to treatment with transforming growth factor beta (TGF beta). We demonstrate in vitro that apoJ protein can be translated from two in-frame ATG sites. Initiation from the first ATG encodes for the secretory form of apoJ and initiation from the second ATG, located 33 amino acids downstream of the first and lacking the hydrophobic signal sequence, encodes for a truncated apoJ protein. This shorter form of apoJ is not recognized by microsomes and therefore not glycosylated, and we postulate that it is retained intracellularly and targeted to the nucleus due to the presence of an SV40-like nuclear localization sequence (NLS). This mechanism of nuclear targeting of apoJ occurs in cells since the protein isolated from nuclei of TGF beta-treated cells and the in vitro-translated truncated form are identical by V8 protease analysis. These results suggest that the diverse physiological responses attributed to apoJ may be elicited through a common molecular mechanism involving a previously uncharacterized intracellular form of the protein.
Reddy
K B
KB
Department of Cell Biology, Cleveland Clinic Research Institute, Ohio 44195-5245, USA.
Jin
G
G
Karode
M C
MC
Harmony
J A
JA
Howe
P H
PH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D051152:Clusterin; D017931:DNA Primers; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008907:Mink; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000378:metabolism*
C495242:CLU protein, human; D051152:Clusterin; D017931:DNA Primers; D006023:Glycoproteins; D018832:Molecular Chaperones; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta