The human ribosomal protein eL29 binds in vivo to the cognate mRNA by interacting with its coding sequence, as revealed from in-cell cross-linking data

Deficiency of the Ribosomal Protein uL5 Leads to Significant Rearrangements of the Transcriptional and Translational Landscapes in Mammalian Cells

“…Given the high expression levels of RPL12, RPLP0 and RPLP1 genes in HEK293T cells (see Table S2), one could conclude that the amounts of excess proteins uL11, uL10 and P1 were quite significant in uL5 knocked down cells, which allowed the proteins to bind to their own coding mRNAs and thereby to reduce the levels of these mRNAs. A similar regulatory feedback mechanism has been found earlier for human genes encoding ribosomal proteins, such as eS26 [25], uS15 [26], eL29 [27] and uS3 [28], which have been shown to be able to bind with their own mRNAs in the cytoplasm or with their pre-mRNAs in the nucleus, inhibiting translation or splicing, respectively. It is quite possible that this mechanism takes place in the regulation of genes encoding uL11, uL10 and P1 as well.…”

“…Rabbit antibodies against GAPDH (Proteintech, #60004-1-Ig) were used as references. One quarter of the lysate (extracted from 5 million cells) was mixed with TRIzol Reagent (Ambion, Waltham, MA, USA) to isolate total cellular RNA, and the remaining three quarters (extracted from 15 million cells) were subjected to sucrose density gradient ultracentrifugation to generate a polysome profile, as described in [27] with minor modifications. Briefly, the extract was layered onto a 5 to 50% linear sucrose gradient in 50 mM buffer Tris-HCl (pH 7.5) containing 100 mM KCl and 12mM MgCl 2 and centrifuged at 19,000 rpm for 17 h at 4 • C in a SW40 rotor.…”