Cell density-dependent nuclear/cytoplasmic localization of NORPEG (RAI14) protein

Journal Article · Fri Jul 14 04:00:00 EDT 2006 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/j.bbrc.2006.04.184· OSTI ID:20854351

Kutty, R Krishnan ^[1]; Chen, Shanyi ^[1]; Samuel, William ^[1]; Vijayasarathy, Camasamudram ^[2]; Duncan, Todd ^[1]; Tsai, Jen-Yue ^[3]; Fariss, Robert N ^[3]; Carper, Deborah ^[4]; Jaworski, Cynthia ^[4]; Wiggert, Barbara ^[1]

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1. Section on Biochemistry, Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, Bethesda, MD 20892 (United States)
2. National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, MD 20892 (United States)
3. Biological Imaging Core, National Eye Institute, National Institutes of Health, Bethesda, MD 20892 (United States)
4. Section on Molecular Therapeutics, National Eye Institute, National Institutes of Health, Bethesda, MD 20892 (United States)

NORPEG (RAI14), a developmentally regulated gene induced by retinoic acid, encodes a 980 amino acid (aa) residue protein containing six ankyrin repeats and a long coiled-coil domain [Kutty et al., J. Biol. Chem. 276 (2001), pp. 2831-2840]. We have expressed aa residues 1-287 of NORPEG and used the recombinant protein to produce an anti-NORPEG polyclonal antibody. Confocal immunofluorescence analysis showed that the subcellular localization of NORPEG in retinal pigment epithelial (ARPE-19) cells varies with cell density, with predominantly nuclear localization in nonconfluent cells, but a cytoplasmic localization, reminiscent of cytoskeleton, in confluent cultures. Interestingly, an evolutionarily conserved putative monopartite nuclear localization signal (P{sup 27}KKRKAP{sup 276}) was identified by analyzing the sequences of NORPEG and its orthologs. GFP-NORPEG (2-287 aa), a fusion protein containing this signal, was indeed localized to nuclei when expressed in ARPE-19 or COS-7 cells. Deletion and mutation analysis indicated that the identified nuclear localization sequence is indispensable for nuclear targeting.

• OSTI ID: 20854351
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 4 Vol. 345; ISSN BBRCA9; ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
AMINO ACIDS
ANTIBODIES
EPITHELIUM
GENES
MICROTUBULES
MUTATIONS
NITRIC OXIDE
RETINOIC ACID
RHODOPSIN