Nuclear localisation of wild type and mutant galectin-3 in transfected cells.
Galectin-3, a member of a family of carbohydrate-binding proteins, is present generally in the cytoplasm of cells. However, galectin 3 can also be located in nuclei under certain conditions although it lacks any known nuclear localisation signal and the mechanism by which the protein is sequestered in nuclei is unknown. Here we describe that Cos-7 cells or rabbit smooth muscle Rb-1 cells transfected with cDNA encoding hamster galectin-3 sequester the protein in nuclei whereas untransfected BHK cells expressing the endogenous hamster lectin or transfected BHK cells over-expressing the protein, do not. Confocal immunofluorescence microscopy of Cos-7 cells or rabbit smooth muscle Rb-1 cells transfected with cDNAs encoding mutants of hamster galectin-3 containing N-terminal or internal deletions shows that nuclear localisation does not require the first 103 amino acid residues of the protein. Further deletion of residues 104-110 dramatically prevents sequestration in nuclei. However, the sequence A104PTGALT110 by itself is not obligatory for nuclear localisation and can be substituted by other unrelated sequences. A truncated galectin-3 protein, that is blocked in nuclear expression, retains carbohydrate-binding activity, making less likely the possibility that severe N-terminal truncations of galectin-3 induce mis-folding leading to aggregation and cytoplasmic sequestration and an incidental effect on nuclear trafficking. These studies indicate that nuclear import and retention of galectin-3 is a property of the CRD domain and is independent of N-terminal domains that others have shown to contain binding domains for various nuclear components.
Gaudin
J C
JC
National Institute for Medical Research, London, UK.
Mehul
B
B
Hughes
R C
RC
D000818:Animals; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D019556:COS Cells; D002467:Cell Nucleus / Q000737:chemistry*; D006224:Cricetinae; D017931:DNA Primers; D018076:DNA, Complementary; D019084:Fluorescent Antibody Technique, Indirect; D037502:Galectin 3; D007668:Kidney / Q000166:cytology; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics; D009130:Muscle, Smooth / Q000166:cytology; D016296:Mutagenesis / Q000502:physiology; D011817:Rabbits; D014162:Transfection; D014404:Tubulin / Q000032:analysis / Q000276:immunology
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D017931:DNA Primers; D018076:DNA, Complementary; D037502:Galectin 3; D008562:Membrane Glycoproteins; D014404:Tubulin