Characterization of nuclear localizing sequences derived from yeast ribosomal protein L29.
Two particular seven-amino-acid segments from yeast ribosomal protein L29 caused a non-nuclear reporter protein to associate almost exclusively with the yeast nucleus. The two L29-derived nuclear localizing sequences were identical in five of the seven residues, many of which were basic amino acids. Generally, localization of the reporter protein was most impaired by replacement of the basic residues. A particular Arg residue was unique; substitution by any amino acid including Lys diminished nuclear localization of the reporter protein. In L29 the corresponding Arg 25----Lys substitution within the nuclear localizing sequence distal to the N-terminus was without effect, as evidence by normal rates of ribosome assembly and cell growth. However, the analogous Arg 8----Lys substitution within the localizing sequence proximal to the N-terminus led to greatly reduced rates of ribosome assembly and cell growth. Finally, when both localizing sequences contained the Arg----Lys substitution a still greater decrease in ribosome assembly and cell growth was observed. These results were as expected if the two short peptide sequences functioned in nuclear localization and/or assembly of yeast ribosomal protein L29.
Underwood
M R
MR
Department of Biochemistry, University of North Carolina, Chapel Hill 27599.
Fried
H M
HM
D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D012322:RNA Precursors / Q000378:metabolism; D012335:RNA, Ribosomal / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000648:ultrastructure; D013329:Structure-Activity Relationship; D001616:beta-Galactosidase / Q000235:genetics
D012322:RNA Precursors; D012335:RNA, Ribosomal; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; C035078:ribosomal protein L29; D001616:beta-Galactosidase