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(a) Figure 10.2
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Computer model of the interaction of a yeast transcriptional factor, GAL4 (blue), and a seventeen-base-pair region of DNA (red). Zinc ions are in yellow. The protein is a dimer; only the DNA recognition region and associated part are shown. Part (b) is a space- lling model of part (a). (Reprinted
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with permission from Nature, 2 April 1992, Vol. 356, p. 411, g. 3b,c. Copyright 1992 Macmillan Magazines Limited.)
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Tamarin: Principles of Genetics, Seventh Edition
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III. Molecular Genetics
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10. Gene Expression: Transcription
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Types of RNA
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TYPES OF RNA
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In the protein synthesis process, three different kinds of RNA serve in three different roles. The rst type is messenger RNA (mRNA), which carries the DNA sequence information to particles in the cytoplasm known as ribosomes, where the messenger RNA is translated. The sec-
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ond type is transfer RNA (tRNA), which brings the amino acids to the ribosomes, where protein synthesis takes place. The third type of RNA is a structural and functional part of the ribosome called ribosomal RNA (rRNA). The general relationship of the roles of these three types of RNA is diagrammed in gure 10.3. In addition, small RNAs play other roles in cellular metabolism, some of which are described later in the chapter.
DNA Transcription
Ribosomal RNA
Transfer RNA
Messenger RNA
+ Ribosomal proteins
+ Amino acid
Modification in eukaryotes
Ribosome
Translation
Growing polypeptide
Next amino acid
Relationship among the three types of RNA ribosomal, transfer, and messenger during protein synthesis. All three types are found together at the ribosome during protein synthesis.
Tamarin: Principles of Genetics, Seventh Edition
III. Molecular Genetics
10. Gene Expression: Transcription
The McGraw Hill Companies, 2001
Ten
Gene Expression: Transcription
We know that DNA does not take part directly in protein synthesis because, in eukaryotes, translation occurs in the cytoplasm, whereas DNA remains in the nucleus. We suspected for a long time that the genetic intermediate in prokaryotes and eukaryotes was RNA because the cytoplasmic RNA concentration increases with increasing protein synthesis, and the cytoplasmic RNAs carry nucleotide sequences complementary to the cell s DNA. Proof of an RNA intermediate came when it was shown that messenger RNA directs protein synthesis.
DNA 5 A T T A A T A T G C C G C G G C T A 3 5 Heat (denature) and add RNA 3 A T A A G C C G T 3 T A T T C G G C A 5 5 RNA A U A A G + C C G U 3 3
PROKARYOTIC DNA TRANSCRIPTION
DNA-RNA Complementarity
What proof do we have that a messenger RNA exists That is, what proof convinced geneticists that gene-sized RNAs (not transfer RNAs or ribosomal RNAs) were found in the cytoplasm that were complementary to the DNA in the nucleus At least two lines of evidence exist. First, it was shown that the RNAs produced by various organisms have base ratios very similar to the base ratios in the same organisms DNA (table 10.1). The second line of evidence comes from experiments by B. Hall, S. Spiegelman, and others using DNA-RNA hybridization. This technique denatures DNA by heating, which causes the two strands of the double helix to separate. When the solution cools, a certain proportion of the DNA strands rejoin and rewind that is, complementary strands nd each other and re-form double helices. When RNA is added to the denatured DNA solution and the solution is cooled slowly, some of the RNA forms double helices with the DNA if the RNA fragments are complementary to a section of the DNA ( g. 10.4). The existence of extensive complementarity between DNA and RNA is a persuasive indication that DNA acts as a template for complementary RNA. In another experiment, DNA-RNA hybridization showed that bacteriophage infection led to the production of phage-speci c messenger RNA. Gene-sized pieces of RNA extracted from Escherichia coli before and after
Cool (renature) 5 A T A A G C C G T 3 Figure 10.4 3 5 A U A A G T A T T C 3
C G C G G C U A 5
DNA-RNA hybridization occurs between DNA and complementary RNA.
Table 10.1 Correspondence of Base Ratios
Between DNA and RNA of the Same Species
RNA %G C E. coli T2 phage Calf thymus gland 52 35 40 DNA %G C 51 35 43
bacteriophage T2 infection were tested to see if they hybridized with the DNA of the T2 phage or with the DNA of the E. coli cell.The RNA in the E. coli cell was found to hybridize with the E. coli DNA before infection but with the T2 DNA after infection. Thus it is apparent that when the phage attacks the E. coli cell, it starts to manufacture RNA complementary to its own DNA and stops the E. coli DNA from serving as a template. Having reached the conclusion that RNA is transcribed (synthesized) from a DNA template and then directs protein synthesis, we look at two questions. First, is this RNA single- or double-stranded Second, is it synthesized (transcribed) from one or both strands of the parental DNA
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