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Nuclear-accumulation kinetics of p9CksHs1 and p9CksHs2 in live plant cells correlate with immunochemical characteristics

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Summary

The two human homologues of the fission yeast cell cycle protein p13suc1 displayed structural characteristics consistent with their existing in solution as differently folded monomers despite 81% identity with respect to their primary structures and both being capable of fulfilling the functions of their homologues in fission and budding yeasts. Carboxyfluorescein-labelled p9CksHs1 and p9CksHs2 retained their native structures. When microinjected into live stamen hair cells ofTradescantia virginiana, the labelled proteins accumulated in the nuclei of the cells. Markedly different nuclearaccumulation kinetics indicated that the human proteins interact differently with other cellular constituents, which supports the proposition that they may have different roles in cellular regulation.

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Abbreviations

Cdk:

cyclin-dependent kinase

tris:

tris(hydroxymethyl)aminomethane

Hepes:

N-(2-hydroxyethyl)piperazine-N′-(3-ethanesulphonic acid)

CF:

5(6)-carboxyfluorescein-N-hydroxysuccinamide ester

SDS-PAGE:

sodium dodecyl sulphatepolyacrylamide gel electrophoresis

IEF:

isoelectric focusing

DEAE:

Sephacel diethylaminoethyl Sephacel

ELISA:

enzyme-linked immunosorbent assay

IgG:

immunoglobulin

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Authors and Affiliations

  1. Lions Cancer Institute, University of Western Australia, Perth, Western Australia

    E. A. Williams & A. C. Carrello

  2. Department of Biology, University of Massachusetts, Amherst, Massachusetts

    P. K. Hepler

  3. Centre for Applied Cancer Studies, University of Western Australia, GPO Box F308, 6001, Perth, WA, Australia

    E. A. Williams & A. C. Carrello

  4. Research School of Biological Sciences, Australian National University, Acton, ACT

    P. C. L. John

Authors
  1. E. A. Williams
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  2. P. K. Hepler
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  3. A. C. Carrello
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  4. P. C. L. John
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Williams, E.A., Hepler, P.K., Carrello, A.C. et al. Nuclear-accumulation kinetics of p9CksHs1 and p9CksHs2 in live plant cells correlate with immunochemical characteristics. Protoplasma 207, 98–105 (1999). https://doi.org/10.1007/BF01294717

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  • DOI: https://doi.org/10.1007/BF01294717

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