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The making of the Fittest: Natural Selection and Adaptation Molecular Genetics of Color Mutations in Rock Pocket Mice Published July 2012 Revised October 2013 www.BioInteractive.org Page 1 of 7 LESSON STUDENT HANDOUT The Making of the Fittest: Natural Selection and Adaptation MOLECULAR GENETICS OF COLOR MUTATIONS IN ROCK POCKET MICE INTRODUCTION THE ROCK POCKET MOUSE The rock pocket mouse, Chaetodipus intermedius, is a small, nocturnal animal found in the deserts of the southwestern United States. Because most rock pocket mice have a sandy, light-colored coat, they are able to blend in with the light color of the desert rocks and sand that they live on. But populations of primarily dark-colored rock pocket mice have been found living in areas where the ground is covered in a dark rock called basalt, which was caused by geologic lava flows thousands of years ago. Scientists have collected data from a population of primarily dark-colored mice living in an area of basalt in Arizona’s Pinacate lava flow, as well as from a nearby light-colored population. Researchers analyzed the data from these two populations to search for the genetic mutation responsible for the dark coat color. Through their analyses, they discovered a mutation in the Mc1r gene that is involved in coat-color determination. THE MC1R GENE The coat color of rock pocket mice is primarily determined by two pigments: eumelanin, which is dark colored, and pheomelanin, which is light colored. The synthesis of these pigments is controlled by the products of several genes, including the Mc1r gene. This gene encodes a protein called melanocortin 1 receptor (MC1R). This receptor is found embedded in the membrane of melanocytes, which are cells specialized for pigment production. The melanocytes of wild-type (nonmutant) mice produce more pheomelanin than eumelanin. The result is a sandy-colored mouse. The mutated version of the Mc1r gene, however, triggers melanocytes to increase the production of eumelanin, resulting in the dark coat-color phenotype. GENE MUTATION A gene mutation is any change in the DNA sequence of a gene. Gene mutations can change the structure of the resulting protein. A change in protein structure can change, negate, or have no effect on function. There are several types of mutations, and several results that mutations can have on the amino acid sequences of proteins. Types of Mutations • Substitution mutation: The replacement of one nucleotide of DNA for another. Mutations that affect a single nucleotide are called point mutations. • Insertion mutation: The addition of one or more nucleotides to the DNA gene sequence. The insertion of nucleotides can result in frameshift mutations. • Deletion mutation: The loss of one or more nucleotides from the DNA gene sequence. The deletion of nucleotides can result in frameshift mutations. Potential Results a Gene Mutation Has on a Protein • Silent mutation: This mutation does not cause a change in the amino acid sequence of the protein; therefore, there is no change in the resulting protein. • Missense mutation: This mutation causes an amino acid in the sequence to be changed to another amino acid. This type of mutation causes a change in the primary structure of the protein (the linear sequence of amino acids), which typically results in a change in the three-dimensional conformation of the protein. • Nonsense mutation: This mutation causes the protein to be truncated (cut short) due to the incorporation of a “stop” signal into the DNA sequence. This results in translation being stopped before the amino acid sequence of the protein is completed. MATERIALS genetic code chart (see page 7 of this handout or a biology textbook) blue, red, and green colored pencils http://www.biointeractive.org/ Molecular Genetics of Color Mutations in Rock Pocket Mice www.BioInteractive.org Page 2 of 7 LESSON STUDENT HANDOUT The Making of the Fittest: Natural Selection and Adaptation PROCEDURE 1. Watch the short film The Making of the Fittest: Natural Selection and Adaptation. 2. Using the DNA nucleotide sequence in the gene tables (page 3), determine the complementary messenger RNA (mRNA) sequence for the portion of the Mc1r gene provided. (Note: You are only transcribing a small portion of the DNA sequence for this protein. The actual gene contains 951 base pairs.) The numbers above some of the columns indicate amino acid positions in the protein sequence. Also indicated is whether the sequence codes for an extracellular, intracellular, or transmembrane part of the protein. 3. Using the mRNA sequence determined in Step 2, determine the resulting amino acid sequence of the MC1R protein. (Note: This is only a portion of the 317 amino acids in the entire protein. The numbers above some of the columns in the tables indicate amino acid positions in the protein sequence.) You may use the genetic code chart provided in your textbook or the one on page 7 of this handout. 4. There are five mutations in the dark-color Mc1r mutant gene. Compare the DNA sequence of the wild-type Mc1r gene with the DNA sequence of the mutant Mc1r gene. Indicate the locations of the five mutations by circling the five single DNA nucleotides that are mutated in the mutant Mc1r gene table. 5. From the introduction, determine whether each of these mutations is a silent, missense, or nonsense mutation. a. Using the mutant Mc1r gene data, shade in the columns (including DNA, mRNA, and amino acid) in the mutant table that contain a silent mutation. Use a blue colored pencil to do this. b. Likewise, use a red colored pencil to shade in the columns that contain a missense mutation. c. Shade any columns that contain nonsense mutations by using a green colored pencil. 6. Answer the questions following the gene tables. http://www.biointeractive.org/ Molecular Genetics of Color Mutations in Rock Pocket Mice www.BioInteractive.org Page 3 of 7 LESSON STUDENT HANDOUT The Making of the Fittest: Natural Selection and Adaptation GENE TABLES WILD-TYPE MC1R GENE (LIGHT-COLORED COAT PHENOTYPE) 015 024 DNA TTG AGG TGG GCG TGT CCG CAA GGA GTG GAG Ex tr ac el lu la r D om ai n I mRNA Amino Acid MUTANT MC1R GENE (DARK-COLORED COAT PHENOTYPE) 015 024 DNA TTG AGG TGG ACG TGT CCG CAA GGA GTG GAG mRNA Amino Acid WILD-TYPE MC1R GENE (LIGHT-COLORED COAT PHENOTYPE) 105 114 DNA CGG GAC CGG TGG GCC CAC TGA CAC CAT GTC Ex tr ac el lu la r D om ai n III mRNA Amino Acid MUTANT MC1R GENE (DARK-COLORED COAT PHENOTYPE) 105 114 DNA CGG GAC CGG TGG ACC CAC TGA CAC CAT GTC mRNA Amino Acid WILD-TYPE MC1R GENE (LIGHT-COLORED COAT PHENOTYPE) 154 163 DNA TCA TAA CAC TGT GAC GGG GCC CGA GCC ACC In tr ac el lu la r D om ai n I mRNA Amino Acid MUTANT MC1R GENE (DARK-COLORED COAT PHENOTYPE) 154 163 DNA TCA TAA CAC TGT GAC GGG ACC CGA GCC ACC mRNA Amino Acid http://www.biointeractive.org/ Molecular Genetics of Color Mutations in Rock Pocket Mice www.BioInteractive.org Page 4 of 7 LESSON STUDENT HANDOUT The Making of the Fittest: Natural Selection and Adaptation WILD-TYPE MC1R GENE (LIGHT-COLORED COAT PHENOTYPE) 208 212 DNA CAC GTG TAC GAA CGT Tr an sm em br an e V mRNA Amino Acid MUTANT MC1R GENE (DARK-COLORED COAT PHENOTYPE) 208 212 DNA CAC GTG TAC GAG CGT mRNA Amino Acid WILD-TYPE MC1R GENE (LIGHT-COLORED COAT PHENOTYPE) 230 239 DNA GAA CAG GTG GTT CCA AAG GCT GAG TTT CCG In tr ac el lu la r D om ai n III mRNA Amino Acid MUTANT MC1R GENE (DARK-COLORED COAT PHENOTYPE) 230 239 DNA GAA CAG GTG GTG CCA AAG GCT GAG TTT CCG mRNA Amino Acid http://www.biointeractive.org/ Molecular Genetics of Color Mutations in Rock Pocket Mice www.BioInteractive