title: Regulation of Metabolism in Mycobacteria and Cyanobacteria

authors: Vickers, Chelsea

subjects: AmtR

abstract: Bacteria encounter changes in their environment and must adapt to these changes in order to survive. Their ability to adapt is determined by their capacity to efficiently regulate their cellular processes. The mechanisms of bacterial regulation at the transcriptional level have been investigated by structural and functional characterisation of the transcription factor AmtR from Mycobacterium smegmatis. M. smegmatis is a soil bacteria capable of utilising alternative nitrogen sources during nitrogen limitation. AmtR is an important transcription factor that regulates the cellular machinery involved in alternative nitrogen metabolism via a novel co-repressor induced mode of regulation. A second pathway that involves post-transcriptional regulation was investigated by preliminary characterisation of a previously unexplored VapBC family of Type II Toxin Antitoxin (TA) systems found in the cyanobacteria Microcystis aeruginosa and Synechocystis sp. PCC6803 (Synechocystis). Investigation of the biochemistry of M. aeruginosa VapBC systems was limited by poor expression, however a single TA system, VapBCMAE43230/20, was successfully purified by incorporating the SUMO fusion protein into the expression construct. VapBMAE43230 and VapCMAE43220 are coexpressed and form a tetrameric complex that appears to be copurified with DNA. Similar to M. aeruginosa, only one system, VapBCslr1209/1210, in Synechocystis was successfully purified and investigated in vitro. Preliminary EMSA assays indicate VapBCslr1209/1210 is autoregulatory and binds to the promoter region of its operon by recognising palindromic sequences. The RNA pentaprobe system demonstrates that VapCslr1210 has metal dependent endoribonuclease activity that can be inhibited by VapBslr1209. Markerless deletion strains of five Synechocystis vapBC operons were made and growth experiments of three of the deletion strains were conducted. Growth experiments identified a reduced growth rate in two of the deletion strains (when compared to the WT strains): ΔvapBCslr1209/1210 and ΔvapBCssl2138/sll1092. Transcriptomic analyses of both deletion strains were conducted. Preliminary data show that both VapBC systems appear to target genes involved in carbon assimilation and metabolism via both photosynthetic and heterotrophic processes. Interestingly, VapBCslr1209/1210 targets an operon involved in glucose transport, which is also the cellular process targeted by the only other well characterised VapBC system from M. smegmatis (VapBC1283/4). These preliminary results point toward a possible similarity in biological function for VapBC systems across the prokaryotic tree.

text: Regulation of Metabolism in Mycobacteria and Cyanobacteria Bacteria encounter changes in their environment and must adapt to these changes in order to survive. Their ability to adapt is determined by their capacity to efficiently regulate their cellular processes. The mechanisms of bacterial regulation at the transcriptional level have been investigated by structural and functional characterisation of the transcription factor AmtR from Mycobacterium smegmatis. M. smegmatis is a soil bacteria capable of utilising alternative nitrogen sources during nitrogen limitation. AmtR is an important transcription factor that regulates the cellular machinery involved in alternative nitrogen metabolism via a novel co-repressor induced mode of regulation. A second pathway that involves post-transcriptional regulation was investigated by preliminary characterisation of a previously unexplored VapBC family of Type II Toxin Antitoxin (TA) systems found in the cyanobacteria Microcystis aeruginosa and Synechocystis sp. PCC6803 (Synechocystis). Investigation of the biochemistry of M. aeruginosa VapBC systems was limited by poor expression, however a single TA system, VapBCMAE43230/20, was successfully purified by incorporating the SUMO fusion protein into the expression construct. VapBMAE43230 and VapCMAE43220 are coexpressed and form a tetrameric complex that appears to be copurified with DNA. Similar to M. aeruginosa, only one system, VapBCslr1209/1210, in Synechocystis was successfully purified and investigated in vitro. Preliminary EMSA assays indicate VapBCslr1209/1210 is autoregulatory and binds to the promoter region of its operon by recognising palindromic sequences. The RNA pentaprobe system demonstrates that VapCslr1210 has metal dependent endoribonuclease activity that can be inhibited by VapBslr1209. Markerless deletion strains of five Synechocystis vapBC operons were made and growth experiments of three of the deletion strains were conducted. Growth experiments identified a reduced growth rate in two of the deletion strains (when compared to the WT strains): ΔvapBCslr1209/1210 and ΔvapBCssl2138/sll1092. Transcriptomic analyses of both deletion strains were conducted. Preliminary data show that both VapBC systems appear to target genes involved in carbon assimilation and metabolism via both photosynthetic and heterotrophic processes. Interestingly, VapBCslr1209/1210 targets an operon involved in glucose transport, which is also the cellular process targeted by the only other well characterised VapBC system from M. smegmatis (VapBC1283/4). These preliminary results point toward a possible similarity in biological function for VapBC systems across the prokaryotic tree.

year: 2015

title: Moonage Daydream: brilliant Bowie film takes big risks to create something truly new

authors: Perrott, Lisa

subjects: music

abstract: Hypnotic, immersive, kaleidoscopic, sublime: Brett Morgen’s film Moonage Daydream has been described as an “experiential cinematic odyssey” and a “colossal tidal wave of vibrant images and overpowering sound”. But as a Bowie fan of 40 years, this film was a transformative experience for me because of the integrity with which Morgen reassembled “the real thing” to make something authentic and new. Inspired by Bowie’s cautionary words about comfort and his philosophy to “always go further into the water than you feel you’re capable of being in”, Morgen embarked on a mission that would take him far outside his comfort zone. Boldly eschewing the conventional biopic format, he immersed himself in a seven-year creative process – one that has led to a subjective but respectful representation of the artist who helped him navigate his own teenage journeys. Propelling the biographical music documentary form beyond the expected conventions of talking heads and expert analysis, Morgen combines a documentary style with music video aesthetics and surrealist assemblage methods to craft a new form. Employing an even more extensive collage approach than he’d used for his Kurt Cobain film Montage of Heck (2015), Morgen treats Moonage Daydream as an audiovisual tapestry, woven from numerous archival materials: songs, vocal recordings, still photographs and film footage derived from music videos, theatrical films, televised interviews and live performance. Morgen brings these things to life by punctuating them with sonic and visual effects. Bravely facing the potential wrath of Bowie devotees, he takes the creative liberty of disassembling and reanimating Bowie’s hand-drawn sketches, storyboards and paintings.

text: Moonage Daydream: brilliant Bowie film takes big risks to create something truly new Hypnotic, immersive, kaleidoscopic, sublime: Brett Morgen’s film Moonage Daydream has been described as an “experiential cinematic odyssey” and a “colossal tidal wave of vibrant images and overpowering sound”. But as a Bowie fan of 40 years, this film was a transformative experience for me because of the integrity with which Morgen reassembled “the real thing” to make something authentic and new. Inspired by Bowie’s cautionary words about comfort and his philosophy to “always go further into the water than you feel you’re capable of being in”, Morgen embarked on a mission that would take him far outside his comfort zone. Boldly eschewing the conventional biopic format, he immersed himself in a seven-year creative process – one that has led to a subjective but respectful representation of the artist who helped him navigate his own teenage journeys. Propelling the biographical music documentary form beyond the expected conventions of talking heads and expert analysis, Morgen combines a documentary style with music video aesthetics and surrealist assemblage methods to craft a new form. Employing an even more extensive collage approach than he’d used for his Kurt Cobain film Montage of Heck (2015), Morgen treats Moonage Daydream as an audiovisual tapestry, woven from numerous archival materials: songs, vocal recordings, still photographs and film footage derived from music videos, theatrical films, televised interviews and live performance. Morgen brings these things to life by punctuating them with sonic and visual effects. Bravely facing the potential wrath of Bowie devotees, he takes the creative liberty of disassembling and reanimating Bowie’s hand-drawn sketches, storyboards and paintings.

year: 2022-09-14

title: An evaluation of the cultural supervision prototype undertaken within the Department of Corrections, Hamilton

authors: Masters-Awatere, Bridgette

subjects: Maori

abstract: Disproportionately high rates of offending and recidivism among Māori are well documented. As part of its attempt to better meet the needs of Māori offenders, the Department of Corrections is developing cultural supervision for staff in the Community Probation Service, the Public Prisons Service and the Psychological Service. The aims of cultural supervision include improving staff members’ knowledge of Māori cultural values, providing support for staff in managing complex cultural issues, and ensuring safe practice and culturally appropriate behaviour. During 2002, the Māori and Psychology Research Unit was contracted to conduct a survey of current practices in relation to cultural supervision and a process evaluation of a prototype of cultural supervision being trialled in the Waikato among probation officers (Hamilton Area) and sentence planners (Waikeria Prison) (Karapu, Masters, Robertson, Trynes, & Waitoki, 2002). Findings from the survey indicated that most staff had informal cultural support or advice available to them. Usually, this was in the form of Māori colleagues within the Department. Less commonly, support was sought from Māori in other organisations, from knowledgeable non-Māori within the Department, kaumātua and kuia, and whānau members. About a third of Corrections staff were receiving some formal cultural supervision, most of whom regarded it positively. Among other staff, both Māori and non-Māori, there was a high level of interest in cultural supervision, and a view that it would be beneficial to their professional practice. Overall, staff felt that the support and advice currently available to them was inadequate. Findings from the evaluation of the prototype suggested that while many of the participants viewed cultural supervision as important for their job, fewer considered that the supervision they were receiving was meeting their needs. Some experienced non-Māori staff seemed to be resistant to the idea of cultural supervision, feeling that they already knew how to relate to Māori offenders. Māori staff were generally enthusiastic about cultural supervision but wanted it to focus on their personal safety as Māori within a “mainstream” institution rather than on their practice. However, both Māori and non-Māori staff generally felt that the cultural supervision sessions provided a safe environment. The prototype appeared to be a good beginning and the model should be stronger when modified in the light of experience.

text: An evaluation of the cultural supervision prototype undertaken within the Department of Corrections, Hamilton Disproportionately high rates of offending and recidivism among Māori are well documented. As part of its attempt to better meet the needs of Māori offenders, the Department of Corrections is developing cultural supervision for staff in the Community Probation Service, the Public Prisons Service and the Psychological Service. The aims of cultural supervision include improving staff members’ knowledge of Māori cultural values, providing support for staff in managing complex cultural issues, and ensuring safe practice and culturally appropriate behaviour. During 2002, the Māori and Psychology Research Unit was contracted to conduct a survey of current practices in relation to cultural supervision and a process evaluation of a prototype of cultural supervision being trialled in the Waikato among probation officers (Hamilton Area) and sentence planners (Waikeria Prison) (Karapu, Masters, Robertson, Trynes, & Waitoki, 2002). Findings from the survey indicated that most staff had informal cultural support or advice available to them. Usually, this was in the form of Māori colleagues within the Department. Less commonly, support was sought from Māori in other organisations, from knowledgeable non-Māori within the Department, kaumātua and kuia, and whānau members. About a third of Corrections staff were receiving some formal cultural supervision, most of whom regarded it positively. Among other staff, both Māori and non-Māori, there was a high level of interest in cultural supervision, and a view that it would be beneficial to their professional practice. Overall, staff felt that the support and advice currently available to them was inadequate. Findings from the evaluation of the prototype suggested that while many of the participants viewed cultural supervision as important for their job, fewer considered that the supervision they were receiving was meeting their needs. Some experienced non-Māori staff seemed to be resistant to the idea of cultural supervision, feeling that they already knew how to relate to Māori offenders. Māori staff were generally enthusiastic about cultural supervision but wanted it to focus on their personal safety as Māori within a “mainstream” institution rather than on their practice. However, both Māori and non-Māori staff generally felt that the cultural supervision sessions provided a safe environment. The prototype appeared to be a good beginning and the model should be stronger when modified in the light of experience.

year: 2003