Identification of a transcript that is down-regulated in senescent human fibroblasts. Cloning, sequence analysis, and regulation of the human L7 ribosomal protein gene.
Normal eukaryotic cells divide only a limited number of times before proliferation ceases due to cellular senescence. We previously reported that a constitutively expressed, non-cell cycle-regulated transcript of unknown identity declines severalfold when human fibroblasts become senescent. We show here, from the sequence of cDNA and genomic clones, that this transcript encodes L7, a structural protein of the large ribosomal subunit. The human L7 protein shares > 90% amino acid identity with the mouse and rat L7 proteins but is shorter than either rodent protein due to fewer basic repetitive motifs at the amino terminus. The position of the first intron is conserved between the mouse and human genes. The L7 mRNA was abundant, stable (t1/2 > 10 h), and polyadenylated in presenescent and senescent human fibroblasts; however, steady state mRNA levels were 5-10-fold lower in senescent cells, whether derived from fetal lung or neonatal foreskin. Quiescent and senescent cells synthesized protein at similar rates, yet only senescent cultures showed a decline in L7 mRNA. The mRNAs encoding five other ribosomal proteins (L5, P1, S3, S6, and S10) behaved similarly. The results suggest that the senescence-associated decline in L7 and other ribosomal protein mRNAs is unrelated to growth state or protein synthetic rate per se and support the view that senescence and quiescence are dissimilar states.
Seshadri
T
T
Department of Cell and Molecular Biology, Lawrence Berkeley Laboratory, University of California, Berkeley 94720.
Uzman
J A
JA
Oshima
J
J
Campisi
J
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D016922:Cellular Senescence; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D003609:Dactinomycin / Q000494:pharmacology; D015536:Down-Regulation; D005347:Fibroblasts / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D008168:Lung / Q000737:chemistry / Q000196:embryology; D008297:Male; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011061:Poly A / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D012333:RNA, Messenger; C053634:RPL7 protein, human; D012269:Ribosomal Proteins; C512845:Rpl7 protein, mouse; D003609:Dactinomycin; D011061:Poly A; D004247:DNA