Assays for ribosomal RNA processing and ribosome assembly.
The synthesis of ribosomes is a major metabolic activity critical for cell growth and homeostasis. Understanding the mechanisms of ribosome biogenesis has important implications for studying both protein synthesis and cell cycle control. This unit describes several techniques for the analysis of rRNA maturation and ribosome assembly adapted for mammalian cells. Metabolic labeling of rRNA and hybridization analysis of precursors can be used to assess changes in rRNA processing that occur under experimental conditions of interest. Separation of preribosomal particles by sucrose gradient centrifugation is suitable for the analysis of proteins associated with preribosomes during their assembly and maturation in the cell nucleus.
Pestov
Dimitri G
DG
University of Medicine and Dentistry of New Jersey, School of Osteopathic Medicine, Department of Cell Biology, Stratford, New Jersey, USA.
Lapik
Yevgeniya R
YR
Lau
Lester F
LF
D000818:Animals; D002466:Cell Nucleolus / Q000737:chemistry / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D003584:Cytological Techniques; D015342:DNA Probes; D006801:Humans; D008322:Mammals / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012335:RNA, Ribosomal / Q000032:analysis / Q000378:metabolism*; D011868:Radioisotopes / Q000032:analysis / Q000378:metabolism; D012261:Ribonucleoproteins / Q000032:analysis / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism*
D015342:DNA Probes; D012335:RNA, Ribosomal; D011868:Radioisotopes; D012261:Ribonucleoproteins