title: DNA repair in Antarctic bacteria - characterisation of two nuclease proteins from Antarctic dry valley metagenomes

authors: Stelzer, Ronja

subjects: Antarctica

abstract: DNA repair processes are crucial for the growth and survival of all organisms. This is especially true for organisms, that inhabit extreme habitats, like the Antarctic Dry Valleys. The Dry Valleys are one of the coldest and driest desert environments on Earth. The conditions in the Dry Valleys, including high levels of UV radiation, low temperatures, multiple daily freeze-thaw cycles, desiccation, and low nutrient levels, are highly damaging to the genomic DNA of the organisms inhabiting them. The Dry Valleys are dominated by microorganisms, which have a range of adaptations to survive in this extreme environment. Previous studies focus largely on the metabolic adaptation of Dry Valley inhabitants. In this thesis, the DNA repair machinery of bacteria in the Dry Valleys was examined to determine how organisms survive the DNA-damaging conditions.
Our understanding of DNA repair and replication in bacteria pathways is largely based on studies of isolated organisms and pathogens, which have been extensively studied. Methods such as metagenomic sequencing allow us to find novel proteins from currently unculturable organisms. In a search for novel DNA repair proteins in bacterial metagenomes from the Dry Valleys, two unique nuclease proteins were chosen for structural and biochemical characterisation. One of these nuclease proteins belongs to the currently uncharacterised protein domain UPF0102. This protein is distantly related to the archaeal Holliday junction resolvases and is a Type II restriction endonuclease type protein. The activity of DV-Hjc and the UPF0102 domain was characterised in vitro using biochemical assays and in vivo in E. coli knockout cells. Biochemical characterisation of the protein showed that the protein has strong binding affinity with double-stranded DNA substrates and Holliday-junction mimicking substrates. The structure of the protein was determined via X-ray crystallography at a resolution of 0.9 Å and structural features were related to its activity, revealing an extensive DNA binding surface and an active site similar to that of archaeal Holliday junction resolvases. The second nuclease characterised in this thesis is part of a unique nuclease-ligase fusion protein and belongs to the nuclease group of MBL-β-CASP proteins. The biochemical characterisation of the protein showed its ability to bind and cleave DNA, specific activity was observed with abasic site substrates.

text: DNA repair in Antarctic bacteria - characterisation of two nuclease proteins from Antarctic dry valley metagenomes DNA repair processes are crucial for the growth and survival of all organisms. This is especially true for organisms, that inhabit extreme habitats, like the Antarctic Dry Valleys. The Dry Valleys are one of the coldest and driest desert environments on Earth. The conditions in the Dry Valleys, including high levels of UV radiation, low temperatures, multiple daily freeze-thaw cycles, desiccation, and low nutrient levels, are highly damaging to the genomic DNA of the organisms inhabiting them. The Dry Valleys are dominated by microorganisms, which have a range of adaptations to survive in this extreme environment. Previous studies focus largely on the metabolic adaptation of Dry Valley inhabitants. In this thesis, the DNA repair machinery of bacteria in the Dry Valleys was examined to determine how organisms survive the DNA-damaging conditions.
Our understanding of DNA repair and replication in bacteria pathways is largely based on studies of isolated organisms and pathogens, which have been extensively studied. Methods such as metagenomic sequencing allow us to find novel proteins from currently unculturable organisms. In a search for novel DNA repair proteins in bacterial metagenomes from the Dry Valleys, two unique nuclease proteins were chosen for structural and biochemical characterisation. One of these nuclease proteins belongs to the currently uncharacterised protein domain UPF0102. This protein is distantly related to the archaeal Holliday junction resolvases and is a Type II restriction endonuclease type protein. The activity of DV-Hjc and the UPF0102 domain was characterised in vitro using biochemical assays and in vivo in E. coli knockout cells. Biochemical characterisation of the protein showed that the protein has strong binding affinity with double-stranded DNA substrates and Holliday-junction mimicking substrates. The structure of the protein was determined via X-ray crystallography at a resolution of 0.9 Å and structural features were related to its activity, revealing an extensive DNA binding surface and an active site similar to that of archaeal Holliday junction resolvases. The second nuclease characterised in this thesis is part of a unique nuclease-ligase fusion protein and belongs to the nuclease group of MBL-β-CASP proteins. The biochemical characterisation of the protein showed its ability to bind and cleave DNA, specific activity was observed with abasic site substrates.

year: 2024-12-09

title: Depression in autobiographical sports writing: A backstage pass into the dark locker room

authors: Shepherd, Caleb

subjects: Depression

abstract: Autobiographical sports writing (ASW) enables authors to provide the general public an insight into their world as an elite athlete. Access to the lives of these athletes is otherwise left to sports media and journalism, which can often skew material for the right headline. This autobiographical insight also allows athletes the opportunity to discuss their battles with depression and mental health without the scrutiny of media and without the risks of giving their competitors an edge. It is important for all autobiographical writers to establish a connection with the reader and recognise that by entering into the field of ASW they are also entering into an unwritten contract that implies their story is truthful. This requires a level of vulnerability and bravery from the writer, particularly with a topic such as depression. The resulting ASW grants the audience a backstage pass into their often dark and confronting ‘locker rooms’. 

This thesis will examine and critically analyse three primary texts from this field: Graeme Obree’s Flying Scotsman, Andre Agassi’s Open and Amanda Beard’s In The Water They Can’t See You Cry. I will undertake a literature-focused consideration of the methods each author uses to craft their narrative. These texts represent individual stories which are unique and set them apart from the vast majority of sports writing. Each author uses specific literary techniques to craft their narratives; which include a combination of first-person narration, present tense, outward expression of internal voice and careful selection of opening scenes. 

This is a creative practice thesis and the critical analysis will be followed by an extract from my own memoir in the same field, The Other Games. The creative component is thematically linked to the critical analysis, examining my personal life as an Olympic rowing coxswain and detailing some of my periodic battles with depression. This section will also incorporate and display some of the literary techniques that bring my chosen texts to life, in particular a carefully selected opening scene and an outward expression of internal voice through italics.

text: Depression in autobiographical sports writing: A backstage pass into the dark locker room Autobiographical sports writing (ASW) enables authors to provide the general public an insight into their world as an elite athlete. Access to the lives of these athletes is otherwise left to sports media and journalism, which can often skew material for the right headline. This autobiographical insight also allows athletes the opportunity to discuss their battles with depression and mental health without the scrutiny of media and without the risks of giving their competitors an edge. It is important for all autobiographical writers to establish a connection with the reader and recognise that by entering into the field of ASW they are also entering into an unwritten contract that implies their story is truthful. This requires a level of vulnerability and bravery from the writer, particularly with a topic such as depression. The resulting ASW grants the audience a backstage pass into their often dark and confronting ‘locker rooms’. 

This thesis will examine and critically analyse three primary texts from this field: Graeme Obree’s Flying Scotsman, Andre Agassi’s Open and Amanda Beard’s In The Water They Can’t See You Cry. I will undertake a literature-focused consideration of the methods each author uses to craft their narrative. These texts represent individual stories which are unique and set them apart from the vast majority of sports writing. Each author uses specific literary techniques to craft their narratives; which include a combination of first-person narration, present tense, outward expression of internal voice and careful selection of opening scenes. 

This is a creative practice thesis and the critical analysis will be followed by an extract from my own memoir in the same field, The Other Games. The creative component is thematically linked to the critical analysis, examining my personal life as an Olympic rowing coxswain and detailing some of my periodic battles with depression. This section will also incorporate and display some of the literary techniques that bring my chosen texts to life, in particular a carefully selected opening scene and an outward expression of internal voice through italics.

year: 2020

title: Mā ngā huruhuru ka rere te manu: Understanding leadership for critical educational reform in Aotearoa New Zealand

authors: Anderson, Zac Kylie Elizabeth

subjects: educational leadership

abstract: The research project presented in this thesis explores the leadership journeys of four first-time principals engaging in Kia Eke Panuku, a professional development initiative in English-medium secondary schools in Aotearoa New Zealand.  It seeks to understand the role these first-time principals play in insitutionalising to depth, educational reform to enable Māori students to pursue their potential as Māori under the Ka Hikitia imperative.  This thesis investigates the type of leadership required to achieve educational reform for Māori learners in Aotearoa, embedded in the inherent promise of the Treaty of Waitangi.
  
This thesis contends these principals contribute to educational reform by bringing criticality to their leadership and embracing an ongoing and iterative process of conscientisation and resistance, leading to transformative praxis.  These principals recognised their role as educators leading pedagogical change in their schools, as well as leading others within their sphere of influence to ultimately change the fabric of society in Aotearoa.  They both learned and led alongside each other, displaying an openness to embracing multiple worldviews with a deeply held moral imperative to eradicate oppression of Māori students.  Such a stance actively creates bicultural spaces within schools, where both Treaty partners can benefit from the principles of the Treaty promises.

text: Mā ngā huruhuru ka rere te manu: Understanding leadership for critical educational reform in Aotearoa New Zealand The research project presented in this thesis explores the leadership journeys of four first-time principals engaging in Kia Eke Panuku, a professional development initiative in English-medium secondary schools in Aotearoa New Zealand.  It seeks to understand the role these first-time principals play in insitutionalising to depth, educational reform to enable Māori students to pursue their potential as Māori under the Ka Hikitia imperative.  This thesis investigates the type of leadership required to achieve educational reform for Māori learners in Aotearoa, embedded in the inherent promise of the Treaty of Waitangi.
  
This thesis contends these principals contribute to educational reform by bringing criticality to their leadership and embracing an ongoing and iterative process of conscientisation and resistance, leading to transformative praxis.  These principals recognised their role as educators leading pedagogical change in their schools, as well as leading others within their sphere of influence to ultimately change the fabric of society in Aotearoa.  They both learned and led alongside each other, displaying an openness to embracing multiple worldviews with a deeply held moral imperative to eradicate oppression of Māori students.  Such a stance actively creates bicultural spaces within schools, where both Treaty partners can benefit from the principles of the Treaty promises.

year: 2018