1. Polar bears are highly adapted for life in cold climates around the North Pole. Brown bears, black bears, and pandas are found in warmer environments. Researchers collected complete mitochondrial DNA sequences from several populations of bears and constructed a phylogenetic tree to represent their evolutionary relatedness (Figure 1).
A researcher studying adaptation in bears sequenced the nuclear gene encoding a lysosomal trafficking protein (LYST) in polar bears, brown bears, black bears, and panda bears. There are seven inferred amino acid substitutions that are found only in polar bears. Mutations that cause similar substitutions in the human LYST protein are associated with Chediak-Higashi syndrome, an autosomal recessive condition in which pigment is absent from the hair and eyes. The researcher used the inferred amino acid sequences to build the distance matrix shown in Table 1.
(a) Use the phylogenetic tree in Figure 1 to estimate the age in hundreds of thousands of years of the most recent common ancestor of all brown bears. Identify the population of brown bears to which polar bears are most closely related based on the mitochondrial DNA sequence comparison. Identify two populations whose positions could be switched without affecting the relationships illustrated in the phylogenetic tree.
(b) Construct a cladogram on the template to represent a model of the evolutionary relatedness among the bear species based on the differences in LYST protein sequences (Table 1). Circle the position on the cladogram that represents the out-group.
(c) A student claims that mitochondrial DNA sequence comparisons provide a more accurate phylogeny of bear species than do LYST protein sequence comparisons. Provide ONE piece of reasoning to support the student’s claim.
(d) A researcher genetically engineers a mouse strain by deleting the mouse lyst gene and replacing it with the polar bear lyst gene. Predict the most likely difference in phenotype of the transgenic mouse strain compared to the wild-type mouse strain. Justify your prediction.
(e) Describe how the mutation in the lyst gene became common in the polar bear population. If the lyst gene were the only determinant of fur color, predict the percent of white offspring produced by a mating between a polar bear and a brown bear.
2. Some pathogenic bacteria enter cells, replicate, and spread to other cells, causing illness in the host organism. Host cells respond to these infections in a number of ways, one of which involves activating particular enzymatic pathways (Figure 1). Cells normally produce a steady supply of inactive caspase-1 protein. In response to intracellular pathogens, the inactive caspase-1 is cleaved and forms an active caspase-1 (step 1). Active caspase-1 can cleave two other proteins. When caspase-1 cleaves an inactive interleukin (step 2), the active portion of the interleukin is released from the cell. An interleukin is a signaling molecule that can activate the immune response. When caspase-1 cleaves gasdermin (step 3), the N-terminal portions of several gasdermin proteins associate in the cell membrane to form large, nonspecific pores.
Researchers created the model in Figure 1 using data from cell fractionation studies. In the experiments, various parts of the cell were separated into fractions by mechanical and chemical methods. Specific proteins known to be located in different parts of the cell were used as markers to determine the location of other proteins. The table below shows the presence of known proteins in specific cellular fractions.
(a) Describe the effect of inhibiting step 3 on the formation of pores AND on the release of interleukin from the cell.
(b) Make a claim about how cleaving inactive caspase-1 results in activation of caspase-1. A student claims that preinfection production of inactive precursors shortens the response time of a cell to a bacterial infection. Provide ONE reason to support the student’s claim.
(c) A student claims that the NF-kB protein is located in the cytoplasm until the protein is needed for transcription. Justify the student’s claim with evidence. Identify TWO fractions where N-terminal gasdermin would be found in cells infected with pathogenic bacteria.
(d) Describe the most likely effect of gasdermin pore formation on water balance in the cell in a hypotonic environment.
(e) Explain how gasdermin pore formation AND interleukin release contribute to an organism’s defense against a bacterial pathogen.