Nuclear targeting of the maize R protein requires two nuclear localization sequences

Journal Article · Mon Feb 01 04:00:00 EST 1993 · Plant Physiology; (United States)

DOI:https://doi.org/10.1104/pp.101.2.353· OSTI ID:6376431

Shieh, M W; Raikhel, N V ^[1]; Wessler, S R ^[2]

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1. Michigan State Univ., East Lansing (United States)
2. 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), Journal Name: Plant Physiology; (United States) Vol. 101:2; ISSN 0032-0889; ISSN PLPHAY
• Country of Publication: United States
• Language: English

Related Subjects

550200 -- Biochemistry
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ARGININE
BIOCHEMISTRY
BIOLOGY
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CEREALS
CHEMISTRY
CYTOPLASM
ENZYMES
GENES
GENETICS
GLUCURONIDASE
GLYCOSYL HYDROLASES
GRAMINEAE
HYDROLASES
LILIOPSIDA
MAGNOLIOPHYTA
MAIZE
MOLECULAR STRUCTURE
O-GLYCOSYL HYDROLASES
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANT CELLS
PLANTS
PROTEIN ENGINEERING
PROTEINS
TRANSCRIPTION
TRANSCRIPTION FACTORS