Actin polymerization and cell motility are affected by NAA80-mediated posttranslational N-terminal acetylation of actin

Aksnes, Henriette; Marie, Michaël; Arnesen, Thomas; Drazic, Adrian

doi:10.1080/19420889.2018.1526572

Cited by 21 publications

(20 citation statements)

References 13 publications

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“…A consistent observation following diploidization of cultures was the tendency for less colony-like tight packing, but rather more separation of cells. We therefore suspected increased random motility of the diploid and conducted a single-cell tracking to characterize the motility properties, similar to previous phenotypic analyses (Aksnes et al, 2018). Indeed, the diploids were here indicated to have an increased motility compared the haploids (Fig.…”

Section: Haploid and Diploid Hap1 Cells Are Fundamentally Differentsupporting

confidence: 74%

Efficient and crucial quality control of HAP1 cell ploidy status

et al. 2020

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The near-haploid human cell line HAP1 recently became a popular subject for CRISPR/Cas9 editing, since only one allele requires modification. Through the gene-editing service at Horizon Discovery, there are at present more than 7500 edited cell lines available and the number continuously increases. The haploid nature of HAP1 is unstable as cultures become diploid with time. Here, we demonstrated some fundamental differences between haploid and diploid HAP1 cells, hence underlining the need for taking control over ploidy status in HAP1 cultures prior to phenotyping. Consequently, we optimized a procedure to determine the ploidy of HAP1 by flow cytometry in order to obtain diploid cultures and avoid ploidy status as an interfering variable in experiments. Furthermore, in order to facilitate this quality control, we validated a size-based cell sorting procedure to obtain the diploid culture more rapidly. Hence, we provide here two streamlined protocols for quality controlling the ploidy of HAP1 cells and document their validity and necessity.This article has an associated First Person interview with the co-first authors of the paper.

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Section: Haploid and Diploid Hap1 Cells Are Fundamentally Differentsupporting

confidence: 74%

Efficient and crucial quality control of HAP1 cell ploidy status

et al. 2020

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“…The 24-well plate was then calibrated with HoloLids for ~ 15 min at 37 °C, 5% CO 2 . Live-cell holographic imaging was set up according to manufacturer's manual as described previously 37 . Three fields of view were monitored at 20 × magnification per well.…”

Section: Methodsmentioning

confidence: 99%

Exploiting the potential of commercial digital holographic microscopy by combining it with 3D matrix cell culture assays

Hellesvik

Øye

Aksnes

2020

Sci Rep

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3D cell culture assays are becoming increasingly popular due to their higher resemblance to tissue environment. These provide an increased complexity compared to the growth on 2D surface and therefore allow studies of advanced cellular properties such as invasion. We report here on the use of 3D Matrigel cell preparations combined with a particular gentle and informative type of live-cell microscopy: quantitative digital holographic microscopy (DHM), here performed by a commercial software-integrated system, currently mostly used for 2D cell culture preparations. By demonstrating this compatibility, we highlight the possible time-efficient quantitative analysis obtained by using a commercial software-integrated DHM system, also for cells in a more advanced 3D culture environment. Further, we demonstrate two very different examples making use of this advantage by performing quantitative DHM analysis of: (1) wound closure cell monolayer Matrigel invasion assay and (2) Matrigel-trapped single and clumps of suspension cells. For both these, we benefited from the autofocus functionality of digital phase holographic imaging to obtain 3D information for cells migrating in a 3D environment. For the latter, we demonstrate that it is possible to quantitatively measure tumourigenic properties like growth of cell clump (or spheroid) over time, as well as single-cell invasion out of cell clump and into the surrounding extracellular matrix. Overall, our findings highlight several possibilities for 3D digital holographic microscopy applications combined with 3D cell preparations, therein studies of drug response or genetic alterations on invasion capacity as well as on tumour growth and metastasis.

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“…Henriette Aksnes (University of Bergen, Norway) presented compelling functional data on N-terminal acetylation of actin, the largest protein modification in cellsnearly 80% of all proteins are N-terminally acetylated. She demonstrated that the N-terminus of both βand γ-actin is acetylated by NAA80, an enzyme whose loss increases lamellipodia and filopodia numbers, causes cells to migrate faster and fragmentation of the Golgi (Aksnes et al, 2018). By using non-migrating NAA80-knockout cells, she convincingly argued that Golgi fragmentation is independent of hypermotility.…”

Section: Actin Diversity and Organelle Movementmentioning

confidence: 99%

Meeting report – Cell dynamics: organelle–cytoskeleton interface

Mogessie

Zenner

Renkawitz³

2019

Journal of Cell Science

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A hallmark of eukaryotic cells is the spatial separation of molecular and biochemical processes into membrane-bound organelles, such as mitochondria, endoplasmic reticulum and Golgi. At the 'Cell dynamics: organelle-cytoskeleton interface' meeting held in Lisbon, researchers from around the world discussed their findings of how the cytoskeleton regulates dynamics, interaction, and function of organelles in health and disease. Organised by Edgar Gomes, Heidi McBride, Sharon Tooze and Michael Way, the meeting created an open, stimulating and collaborative environment for scientific exchange and an opportunity to highlight the newest trends in the field.

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Actin polymerization and cell motility are affected by NAA80-mediated posttranslational N-terminal acetylation of actin

Cited by 21 publications

References 13 publications

Efficient and crucial quality control of HAP1 cell ploidy status

Efficient and crucial quality control of HAP1 cell ploidy status

Exploiting the potential of commercial digital holographic microscopy by combining it with 3D matrix cell culture assays

Meeting report – Cell dynamics: organelle–cytoskeleton interface

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