Effect of Mine Runoff Metals on the Cyclin-Dependent Kinase 3 (CDK3) Gene of Tetrahymena thermophila

Abstract

Heavy metal contamination in Montana streams via mine runoff represents a significant environmental and public health hazard. Butte’s Berkeley Pit, which contains high levels of copper, iron, arsenic, cadmium, zinc, and sulfuric acid, is at risk of contaminating the groundwater of the Butte valley in the near future if measures are not taken to keep its water level from reaching a critical point.1 Heavy metal toxicity in humans results, sometimes irreparably, in neurological and muscular disease.2 This research investigated how heavy metals from mine runoff affected the growth rate and expression of the Cyclin-dependent Kinase-3 (CDK3) gene in Tetrahymena thermophila. It was hypothesized that exposure to the runoff metals would result in a significant change in the expression of (CDK3) and a significant change in the growth rate of T. thermophila cultures. CDK3 is highly expressed in the early stages of conjugation during meiosis initiation.3 CDK3 complexes with cyclin-dependent kinases and conjugation-specific cyclins to initiate meiosis, however, when a CDK3 deletion occurs, conjugation is arrested at the point of pair formation, therefore it is an essential enzyme required for meiotic division.3 T. thermophila were cultured in a nutrient-poor NEFF media in two separate trial periods, each containing two control and two experimental samples. Experimental cultures were exposed to a solution of simulated mine runoff and analyzed every 7 days. Growth curves were generated during the exposure period by counting the cells every 24 hours using a hemocytometer. Following the 7-day treatment period, RNA was extracted from both control and test samples. Quantitative Polymerase Chain Reaction (qPCR) was used to analyze CDK3 expression in response to the treatment. It was predicted there would be a significant change in CDK3 gene expression due to heavy metal exposure, as well as decreased conjugation rates in comparison to control cultures.Heavy metal contamination in Montana streams via mine runoff represents a significant environmental and public health hazard. Butte’s Berkeley Pit, which contains high levels of copper, iron, arsenic, cadmium, zinc, and sulfuric acid, is at risk of contaminating the groundwater of the Butte valley in the near future if measures are not taken to keep its water level from reaching a critical point.1 Heavy metal toxicity in humans results, sometimes irreparably, in neurological and muscular disease.2 This research investigated how heavy metals from mine runoff affected the growth rate and expression of the Cyclin-dependent Kinase-3 (CDK3) gene in Tetrahymena thermophila. It was hypothesized that exposure to the runoff metals would result in a significant change in the expression of (CDK3) and a significant change in the growth rate of T. thermophila cultures. CDK3 is highly expressed in the early stages of conjugation during meiosis initiation.3 CDK3 complexes with cyclin-dependent kinases and conjugation-specific cyclins to initiate meiosis, however, when a CDK3 deletion occurs, conjugation is arrested at the point of pair formation, therefore it is an essential enzyme required for meiotic division.3 T. thermophila were cultured in a nutrient-poor NEFF media in two separate trial periods, each containing two control and two experimental samples. Experimental cultures were exposed to a solution of simulated mine runoff and analyzed every 7 days. Growth curves were generated during the exposure period by counting the cells every 24 hours using a hemocytometer. Following the 7-day treatment period, RNA was extracted from both control and test samples. Quantitative Polymerase Chain Reaction (qPCR) was used to analyze CDK3 expression in response to the treatment. It was predicted there would be a significant change in CDK3 gene expression due to heavy metal exposure, as well as decreased conjugation rates in comparison to control cultures.Heavy metal contamination in Montana streams via mine runoff represents a significant environmental and public health hazard. Butte’s Berkeley Pit, which contains high levels of copper, iron, arsenic, cadmium, zinc, and sulfuric acid, is at risk of contaminating the groundwater of the Butte valley in the near future if measures are not taken to keep its water level from reaching a critical point.1 Heavy metal toxicity in humans results, sometimes irreparably, in neurological and muscular disease.2 This research investigated how heavy metals from mine runoff affected the growth rate and expression of the Cyclin-dependent Kinase-3 (CDK3) gene in Tetrahymena thermophila. It was hypothesized that exposure to the runoff metals would result in a significant change in the expression of (CDK3) and a significant change in the growth rate of T. thermophila cultures. CDK3 is highly expressed in the early stages of conjugation during meiosis initiation.3 CDK3 complexes with cyclin-dependent kinases and conjugation-specific cyclins to initiate meiosis, however, when a CDK3 deletion occurs, conjugation is arrested at the point of pair formation, therefore it is an essential enzyme required for meiotic division.3 T. thermophila were cultured in a nutrient-poor NEFF media in two separate trial periods, each containing two control and two experimental samples. Experimental cultures were exposed to a solution of simulated mine runoff and analyzed every 7 days. Growth curves were generated during the exposure period by counting the cells every 24 hours using a hemocytometer. Following the 7-day treatment period, RNA was extracted from both control and test samples. Quantitative Polymerase Chain Reaction (qPCR) was used to analyze CDK3 expression in response to the treatment. It was predicted there would be a significant change in CDK3 gene expression due to heavy metal exposure, as well as decreased conjugation rates in comparison to control cultures.Heavy metal contamination in Montana streams via mine runoff represents a significant environmental and public health hazard. Butte’s Berkeley Pit, which contains high levels of copper, iron, arsenic, cadmium, zinc, and sulfuric acid, is at risk of contaminating the groundwater of the Butte valley in the near future if measures are not taken to keep its water level from reaching a critical point.1 Heavy metal toxicity in humans results, sometimes irreparably, in neurological and muscular disease.2 This research investigated how heavy metals from mine runoff affected the growth rate and expression of the Cyclin-dependent Kinase-3 (CDK3) gene in Tetrahymena thermophila. It was hypothesized that exposure to the runoff metals would result in a significant change in the expression of (CDK3) and a significant change in the growth rate of T. thermophila cultures. CDK3 is highly expressed in the early stages of conjugation during meiosis initiation.3 CDK3 complexes with cyclin-dependent kinases and conjugation-specific cyclins to initiate meiosis, however, when a CDK3 deletion occurs, conjugation is arrested at the point of pair formation, therefore it is an essential enzyme required for meiotic division.3 T. thermophila were cultured in a nutrient-poor NEFF media in two separate trial periods, each containing two control and two experimental samples. Experimental cultures were exposed to a solution of simulated mine runoff and analyzed every 7 days. Growth curves were generated during the exposure period by counting the cells every 24 hours using a hemocytometer. Following the 7-day treatment period, RNA was extracted from both control and test samples. Quantitative Polymerase Chain Reaction (qPCR) was used to analyze CDK3 expression in response to the treatment. It was predicted there would be a significant change in CDK3 gene expression due to heavy metal exposure, as well as decreased conjugation rates in comparison to control cultures.Heavy metal contamination in Montana streams via mine runoff represents a significant environmental and public health hazard. Butte’s Berkeley Pit, which contains high levels of copper, iron, arsenic, cadmium, zinc, and sulfuric acid, is at risk of contaminating the groundwater of the Butte valley in the near future if measures are not taken to keep its water level from reaching a critical point.1 Heavy metal toxicity in humans results, sometimes irreparably, in neurological and muscular disease.2 This research investigated how heavy metals from mine runoff affected the growth rate and expression of the Cyclin-dependent Kinase-3 (CDK3) gene in Tetrahymena thermophila. It was hypothesized that exposure to the runoff metals would result in a significant change in the expression of (CDK3) and a significant change in the growth rate of T. thermophila cultures. CDK3 is highly expressed in the early stages of conjugation during meiosis initiation.3 CDK3 complexes with cyclin-dependent kinases and conjugation-specific cyclins to initiate meiosis, however, when a CDK3 deletion occurs, conjugation is arrested at the point of pair formation, therefore it is an essential enzyme required for meiotic division.3 T. thermophila were cultured in a nutrient-poor NEFF media in two separate trial periods, each containing two control and two experimental samples. Experimental cultures were exposed to a solution of simulated mine runoff and analyzed every 7 days. Growth curves were generated during the exposure period by counting the cells every 24 hours using a hemocytometer. Following the 7-day treatment period, RNA was extracted from both control and test samples. Quantitative Polymerase Chain Reaction (qPCR) was used to analyze CDK3 expression in response to the treatment. It was predicted there would be a significant change in CDK3 gene expression due to heavy metal exposure, as well as decreased conjugation rates in comparison to control cultures.