Isolation and characterization of a carrot nucleolar protein with structural and sequence similarity to the vertebrate PESCADILLO protein
- Authors
- Ueda, K., Xu, Z.J., Miyagi, N., Ono, M., Wabiko, H., Masuda, K., and Inoue, M.
- ID
- ZDB-PUB-130611-26
- Date
- 2013
- Source
- Plant science : an international journal of experimental plant biology 208: 83-92 (Journal)
- Registered Authors
- Keywords
- Daucus carota, green fluorescent protein (GFP), nuclear localization signal (NLS), nuclear matrix, nucleolus, PESCADILLO
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Cell Nucleolus/metabolism
- DNA, Complementary/isolation & purification
- Daucus carota/cytology
- Daucus carota/genetics
- Daucus carota/metabolism*
- Gene Expression Regulation, Plant
- Humans
- Molecular Sequence Data
- Nicotiana/cytology
- Nuclear Matrix/metabolism
- Nuclear Proteins/chemistry*
- Nuclear Proteins/genetics
- Nuclear Proteins/isolation & purification
- Nuclear Proteins/metabolism
- Plant Proteins/chemistry*
- Plant Proteins/genetics
- Plant Proteins/isolation & purification*
- Plant Proteins/metabolism
- Protein Structure, Tertiary
- Protein Transport
- Sequence Homology, Amino Acid*
- Structural Homology, Protein*
- Subcellular Fractions/metabolism
- Vertebrates/metabolism*
- Zebrafish
- PubMed
- 23683933 Full text @ Plant Sci.
The nuclear matrix is involved in many nuclear events, but its protein architecture in plants is still not fully understood. A cDNA clone was isolated by immunoscreening with a monoclonal antibody raised against nuclear matrix proteins of Daucus carota L. Its deduced amino acid sequence showed about 40% identity with the PESCADILLO protein of zebrafish and humans. Primary structure analysis of the protein revealed a Pescadillo N-terminus domain, a single breast cancer C-terminal domain, two nuclear localization signals, and a potential coiled-coil region as also found in animal PESCADILLO proteins. Therefore, we designated this gene DcPES1. Although DcPES1 mRNA was detected in all tissues examined, its levels were highest in tissues with proliferating cells. Immunofluorescence using specific antiserum against the recombinant protein revealed that DcPES1 localized exclusively in the nucleolus. Examination of fusion proteins with green fluorescent protein revealed that the N-terminal portion was important for localization to the nucleoli of tobacco and onion cells. Moreover, when the nuclear matrix of carrot cells was immunostained with an anti-DcPES1 serum, the signal was detected in the nucleolus. Therefore, the DcPES1 protein appears to be a component of or tightly bound to components of the nuclear matrix.