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 rRN...

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Published inCurrent protocols in cell biology Vol. Chapter 22; p. Unit 22.11
Main Authors Pestov, Dimitri G, Lapik, Yevgeniya R, Lau, Lester F
Format Journal Article
LanguageEnglish
Published United States 01.06.2008
Subjects
Animals
Cell Nucleolus - chemistry
Cell Nucleolus - metabolism
Centrifugation, Density Gradient
Cytological Techniques
DNA Probes
Humans
Mammals - metabolism
Radioisotopes - analysis
Radioisotopes - metabolism
Ribonucleoproteins - analysis
Ribonucleoproteins - metabolism
Ribosomes - chemistry
Ribosomes - metabolism
RNA Processing, Post-Transcriptional
RNA, Ribosomal - analysis
RNA, Ribosomal - metabolism
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Summary: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.
ISSN:1934-2616
DOI:10.1002/0471143030.cb2211s39