Nuclear targeting of the maize R protein requires two nuclear localization sequences
- Michigan State Univ., East Lansing (United States)
- Univ. of Georgia, Athens (United States)
Previous genetic and structural evidence indicates that the maize R gene encodes a nuclear transcriptional activating factor. In-frame carboxyl- and amino-terminal fusions of the R gene to the reporter gene encoding [beta]-glucuronidase (GUS) were sufficient to direct GUS to the nucleus of the transiently transformed onion (Allium cepa) epidermal cells. Further analysis of chimeric constructs containing regions of the R gene fused to the GUS cDNA revealed three specific nuclear localization sequences (NLSs) that were capable of redirecting the GUS protein to the nucleus. Amino-terminal NLS-A (amino acids 100-109, GDRRAAPARP) contained several arginine residues; a similar localization signal is found in only a few viral proteins. The medial NLS-M (amino acids 419-428, MSERKRREKL) is a simian virus 40 large T antigen-type NLS, and the carboxyl-terminal NLS-C (amino acids 598-610, MISESLRKAIGKR) is a mating type [alpha]2 type. NLSs M and C are independently sufficient to direct the GUS protein to the nucleus when it is fused at the amino terminus of GUS, whereas NLS-A fused to GUS partitioned between the nucleus and cytoplasm. Similar partitioning was observed when localization signals NLS-A and NLS-C were independently fused to the carboxy-terminal portion of GUS. A sequential deletion of the localization signals indicated that the amino-terminal and carboxyl-terminal fusions of R and GUS were redirected to the nucleus only when both NLS-A and -M, or NLS-C and -M, were present. These results indicate that multiple localization signals are necessary for nuclear targeting of this protein. The conservation of the localization signals within the alleles of R and similar proteins from other organisms is also discussed. 45 refs., 6 figs.
- OSTI ID:
- 6376431
- Journal Information:
- Plant Physiology; (United States), Vol. 101:2; ISSN 0032-0889
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
GENES
MOLECULAR STRUCTURE
PROTEINS
AMINO ACIDS
ARGININE
BIOCHEMISTRY
CYTOPLASM
GENETICS
GLUCURONIDASE
MAIZE
PLANT CELLS
PROTEIN ENGINEERING
TRANSCRIPTION
TRANSCRIPTION FACTORS
BIOLOGY
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CEREALS
CHEMISTRY
ENZYMES
GLYCOSYL HYDROLASES
GRAMINEAE
HYDROLASES
LILIOPSIDA
MAGNOLIOPHYTA
O-GLYCOSYL HYDROLASES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANTS
550400* - Genetics
550200 - Biochemistry