Conserved Sr Protein Kinase Functions in Nuclear Import and Its Action Is Counteracted by Arginine Methylation in Saccharomyces cerevisiae

doi:10.1083/jcb.150.4.707

Published online 21 August 2000. doi:10.1083/jcb.150.4.707

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© The Rockefeller University Press, 0021-9525/2000//707 $5.00
The Journal of Cell Biology, Volume 150, Number 4, , 2000 707-718

Original Article

Conserved Sr Protein Kinase Functions in Nuclear Import and Its Action Is Counteracted by Arginine Methylation in Saccharomyces cerevisiae

Chi Y. Yun^a and Xiang-Dong Fu^a

^a Department of Cellular and Molecular Medicine, University of California at San Diego, La Jolla, California 92093-0651
Department of Cellular and Molecular Medicine, University of California at San Diego, 9500 Gilman Dr., La Jolla, CA 92093-0651.(858) 534-8549(858) 534-4937

xdfu{at}ucsd.edu

Mammalian serine and arginine–rich (SR) proteins playimportant roles in both constitutive and regulated splicing,and SR protein–specific kinases (SRPKs) are conservedfrom humans to yeast. Here, we demonstrate a novel functionof the single conserved SR protein kinase Sky1p in nuclear importin budding yeast. The yeast SR-like protein Npl3p is known toenter the nucleus through a composite nuclear localization signal(NLS) consisting of a repetitive arginine- glycine-glycine (RGG)motif and a nonrepetitive sequence. We found that the latteris the site for phosphorylation by Sky1p and that this phosphorylationregulates nuclear import of Npl3p by modulating the interactionof the RGG motif with its nuclear import receptor Mtr10p. TheRGG motif is also methylated on arginine residues, but methylationdoes not affect the Npl3p–Mtr10p interaction in vitro.Remarkably, arginine methylation interferes with Sky1p-mediatedphosphorylation, thereby indirectly influencing the Npl3p–Mtr10pinteraction in vivo and negatively regulating nuclear importof Npl3p. These results suggest that nuclear import of Npl3pis coordinately influenced by methylation and phosphorylationin budding yeast, which may represent conserved components inthe dynamic regulation of RNA processing in higher eukaryoticcells.

Key Words: serine and arginine–rich protein kinase • RNA binding protein • nuclear transport • phosphorylation • arginine methylation

Abbreviations used in this paper: GFP, green fluorescent protein;GST, glutathione S-transferase; hnRNP, heterogeneous nuclearribonucleoprotein; NLS, nuclear localization signal; PrA, proteinA; RGG, arginine-glycine-glycine; RRM, RNA recognition motif;RS, arginine- and serine-rich; SAH, S-adenosyl-L-homocysteine;SAM, S-adenosyl-L-methionine; SR, serine and arginine–rich;SRPK, SR protein–specific kinase.

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