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Carboxyl-terminal basic amino acids in the X domain are essential for the nuclear import of phospholipase C delta1.
- Source :
-
Genes to cells : devoted to molecular & cellular mechanisms [Genes Cells] 2002 Sep; Vol. 7 (9), pp. 985-96. - Publication Year :
- 2002
-
Abstract
- Background: Although phospholipase C (PLC)delta1 containing a functional nuclear export signal (NES) is normally localized at the plasma membrane and in the cytoplasm, it shuttles between the nucleus and the cytoplasm. Since nucleocytoplasmic shuttling of a molecule is generally regulated by a balance between its NES and the nuclear localization signal (NLS), we examined whether PLCdelta1 contains an NLS sequence.<br />Results: A region corresponding to the C terminus of the X domain and the XY-linker, which contains clusters of basic amino acid residues, was essential for the nuclear import of PLCdelta1 in Madin-Darby canine kidney cells. A series of point mutations on lysine residues in this region revealed that K432 and K434 in combination were important for the nuclear import. A short synthetic peptide corresponding to residues 429-442, however, was not able to function as an NLS sequence when they were injected into the cytoplasm in a carrier-conjugated form. Neither a longer peptide equivalent to PLCdelta1 412-498 fused to a protein tag consisting of glutathione S-transferase and green fluorescent protein was imported to the nucleus after microinjection into the cytoplasm.<br />Conclusion: The nuclear import of PLCdelta1 requires the C-terminus of the X domain, particularly the amino acid residues K432 and K434, and the XY-linker. The region alone, however, cannot serve as a functional NLS. The machinery for nuclear transport may require additional structural component(s) of the enzyme.
- Subjects :
- Amino Acid Sequence
Animals
Cell Line
Dogs
Green Fluorescent Proteins
Isoenzymes chemistry
Isoenzymes genetics
Kidney cytology
Luminescent Proteins metabolism
Lysine metabolism
Microinjections
Models, Biological
Models, Molecular
Molecular Sequence Data
Nuclear Localization Signals
Peptides genetics
Peptides metabolism
Phospholipase C delta
Point Mutation
Protein Structure, Tertiary
Recombinant Fusion Proteins metabolism
Sequence Alignment
Type C Phospholipases chemistry
Type C Phospholipases genetics
Active Transport, Cell Nucleus physiology
Isoenzymes metabolism
Type C Phospholipases metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1356-9597
- Volume :
- 7
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Genes to cells : devoted to molecular & cellular mechanisms
- Publication Type :
- Academic Journal
- Accession number :
- 12296828
- Full Text :
- https://doi.org/10.1046/j.1365-2443.2002.00577.x