Characterization, subcellular localization and nuclear targeting of casein kinase 2 from Zea mays.

Abstract:

:We have isolated and characterized the genomic clone of maize casein kinase 2 (CK2) alpha subunit using the previously described alphaCK2-1 cDNA clone as a probe. The genomic clone is 7.5 kb long and contains 10 exons, separated by 9 introns of different size, two larger than 1.5 kb and the others around 100-150 bp. The sequence of the exons is 100% homologous to the sequence of the alphaCK2-1 cDNA. Southern hybridization of total genomic DNA from maize embryos with aCK2 cDNA indicated that the alphaCK2-1 gene is part of a multigenic family. We also isolated a new embryo cDNA clone coding for an alphaCK2-2 subunit. We studied the regulation of the enzyme in embryos at the mRNA level, at the protein level and by activity testing. By using immunocytochemistry the CK2 protein was localized in several types of cells of mature embryos. Particularly strong signals were visible in the cytoplasm of epidermis and meristematic cells. Decoration of nuclei of root cortex and scutellum cells was also observed suggesting that CK2 can shift from the cytoplasm into nuclei in specific cell types. We examined whether CK2 contained specific protein domains which actively target the protein to the nucleus by using in-frame fusions of the maize CK2alpha subunit to the reporter gene encoding beta-glucuronidase (GUS) which were assayed in transiently transformed onion epidermal cells. Analysis of chimeric constructs identified one region containing a nuclear localization signal (NLS) that is highly conserved in other alphaCK2 proteins.

journal_name

Plant Mol Biol

journal_title

Plant molecular biology

authors

Peracchia G,Jensen AB,Culiáñez-Macià FA,Grosset J,Goday A,Issinger OG,Pagès M

doi

10.1023/a:1006196530079

subject

Has Abstract

pub_date

1999-05-01 00:00:00

pages

199-211

issue

2

eissn

0167-4412

issn

1573-5028

journal_volume

40

pub_type

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