Extraribosomal functions associated with the C terminus of the 37/67 kDa laminin receptor are required for maintaining cell viability.
The 37/67 kDa laminin receptor (LAMR) is a multifunctional protein, acting as an extracellular receptor, localizing to the nucleus, and playing roles in rRNA processing and ribosome assembly. LAMR is important for cell viability; however, it is unclear which of its functions are essential. We developed a silent mutant LAMR construct, resistant to siRNA, to rescue the phenotypic effects of knocking down endogenous LAMR, which include inhibition of protein synthesis, cell cycle arrest, and apoptosis. In addition, we generated a C-terminal-truncated silent mutant LAMR construct structurally homologous to the Archaeoglobus fulgidus S2 ribosomal protein and missing the C-terminal 75 residues of LAMR, which displays more sequence divergence. We found that HT1080 cells stably expressing either silent mutant LAMR construct still undergo arrest in the G1 phase of the cell cycle when treated with siRNA. However, the expression of full-length silent mutant LAMR rescues cell viability, whereas the expression of the C-terminal-truncated LAMR does not. Interestingly, we also found that both silent mutant constructs restore protein translation and localize to the nucleus. Our findings indicate that the ability of LAMR to regulate viability is associated with its C-terminal 75 residues. Furthermore, this function is distinct from its role in cell proliferation, independent of its ribosomal functions, and may be regulated by a nonnuclear localization.
NYU Cancer Institute, NYU Gene Therapy Center, NYU School of Medicine, New York, NY 10016, USA.
J
J
Scheiman
K V
KV
Jamieson
J
J
Ziello
J-C
JC
Tseng
D
D
Meruelo
D000595:Amino Acid Sequence; D000596:Amino Acids; D001483:Base Sequence; D045744:Cell Line, Tumor; D002467:Cell Nucleus; D002470:Cell Survival; D016193:G1 Phase; D055785:Gene Knockdown Techniques; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000378:metabolism; D017474:Receptors, Laminin / Q000737:chemistry* / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D013329:Structure-Activity Relationship
D000596:Amino Acids; D034741:RNA, Small Interfering; C510624:RPSA protein, human; D017474:Receptors, Laminin; D012269:Ribosomal Proteins
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