JNK2 controls fragmentation of the Golgi complex and the G2/M transition through phosphorylation of GRASP65

Cervigni, Romina Inès; Bonavita, Raffaella; Barretta, Maria Luisa; Spano, Daniela; Ayala, Inmaculada; Nakamura, Nobuhiro; Corda, Daniela; Colanzi, Antonino

doi:10.1242/jcs.164871

Cited by 52 publications

(99 citation statements)

References 50 publications

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“…The initial fragmentation is an important checkpoint for progression through G2/M, and the eventual dispersal of Golgi into small cytosolic vesicles is believed to enable equal partitioning of Golgi elements between daughter cells [52, 55–57]. We initially hypothesized that these dynamic changes in Golgi morphology might be a trigger for L2 translocation.…”

Section: Discussionmentioning

confidence: 99%

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

Calton¹,

Bronnimann²,

Manson³

et al. 2017

PLoS Pathog

114

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The human papillomavirus type 16 (HPV16) L2 protein acts as a chaperone to ensure that the viral genome (vDNA) traffics from endosomes to the trans-Golgi network (TGN) and eventually the nucleus, where HPV replication occurs. En route to the nucleus, the L2/vDNA complex must translocate across limiting intracellular membranes. The details of this critical process remain poorly characterized. We have developed a system based on subcellular compartmentalization of the enzyme BirA and its cognate substrate to detect membrane translocation of L2-BirA from incoming virions. We find that L2 translocation requires transport to the TGN and is strictly dependent on entry into mitosis, coinciding with mitotic entry in synchronized cells. Cell cycle arrest causes retention of L2/vDNA at the TGN; only release and progression past G2/M enables translocation across the limiting membrane and subsequent infection. Microscopy of EdU-labeled vDNA reveals a rapid and dramatic shift in vDNA localization during early mitosis. At late G2/early prophase vDNA egresses from the TGN to a pericentriolar location, accumulating there through prometaphase where it begins to associate with condensed chromosomes. By metaphase and throughout anaphase the vDNA is seen bound to the mitotic chromosomes, ensuring distribution into both daughter nuclei. Mutations in a newly defined chromatin binding region of L2 potently blocked translocation, suggesting that translocation is dependent on chromatin binding during prometaphase. This represents the first time a virus has been shown to functionally couple the penetration of limiting membranes to cellular mitosis, explaining in part the tropism of HPV for mitotic basal keratinocytes.

show abstract

Section: Discussionmentioning

confidence: 99%

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

Calton¹,

Bronnimann²,

Manson³

et al. 2017

PLoS Pathog

114

View full text Add to dashboard Cite

show abstract

“…Golgi fragmentation occurs in a variety of physiological and pathological conditions, and it is often used as readout for various biological processes (e.g mitosis, viral infection, neurodegenerative diseases) 58–60 . Accurate quantification of this phenomenon is therefore essential in studying numerous biological questions.…”

Section: Discussionmentioning

confidence: 99%

High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry

Wortzel

Koifman

Rotter

et al. 2017

Sci Rep

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The Golgi apparatus is a dynamic organelle, which regulates the vesicular trafficking. While cellular trafficking requires active changes of the Golgi membranes, these are not accompanied by changes in the general Golgi’s structure. However, cellular processes such as mitosis, apoptosis and migration require fragmentation of the Golgi complex. Currently, these changes are most commonly studied by basic immunofluorescence and quantified by manual and subjective classification of the Golgi structure in 100–500 stained cells. Several other high-throughput methods exist as well, but those are either complicated or do not provide enough morphological information. Therefore, a simple and informative high content methodology should be beneficial for the study of Golgi architecture. Here we describe the use of high-throughput imaging flow cytometry for quantification of Golgi fragmentation, which provides a simple way to analyze the changes in an automated, quantitative and non-biased manner. Furthermore, it provides a rapid and accurate way to analyze more than 50,000 cells per sample. Our results demonstrate that this method is robust and statistically powerful, thus, providing a much-needed analytical tool for future studies on Golgi dynamics, and can be adapted to other experimental systems.

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“…Moreover, here we show that ERK1c translocates to the Golgi only at prophase, and therefore, it is unlikely to execute the GRASP phosphorylation, which has already started at G2. Instead, it is possible that GRASP proteins are phosphorylated by the other MAPK family member, JNK2, which has recently been shown to mediate GRASP65 phosphorylation at G2 (Cervigni et al, 2015). The substrates of Fig.…”

Section: Discussionmentioning

confidence: 99%

Mitotic Golgi translocation of ERK1c is mediated by PI4KIIIβ/14-3-3γ shuttling complex

Wortzel

Hanoch

Porat

et al. 2015

Journal of Cell Science

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Golgi fragmentation is a highly regulated process that allows division of the Golgi complex between the two daughter cells. The mitotic reorganization of the Golgi is accompanied by a temporary block in Golgi functioning, as protein transport in and out of the Golgi stops. Our group has previously demonstrated the involvement of the alternatively spliced variants ERK1c and MEK1b (ERK1 is also known as MAPK3, and MEK1 as MAP2K1) in mitotic Golgi fragmentation. We had also found that ERK1c translocates to the Golgi at the G2 to M phase transition, but the molecular mechanism underlying this recruitment remains unknown. In this study, we narrowed the translocation timing to prophase and prometaphase, and elucidated its molecular mechanism. We found that CDK1 phosphorylates Ser343 of ERK1c, thereby allowing the binding of phosphorylated ERK1c to a complex that consists of PI4KIIIβ (also known as PI4KB) and the 14-3-3γ dimer (encoded by YWHAB). The stability of the complex is regulated by protein kinase D (PKD)-mediated phosphorylation of PI4KIIIβ. The complex assembly induces the Golgi shuttling of ERK1c, where it is activated by MEK1b, and induces Golgi fragmentation. Our work shows that protein shuttling to the Golgi is not completely abolished at the G2 to M phase transition, thus integrating several independent Golgiregulating processes into one coherent pathway.

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JNK2 controls fragmentation of the Golgi complex and the G2/M transition through phosphorylation of GRASP65

Cited by 52 publications

References 50 publications

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis

High Throughput Analysis of Golgi Structure by Imaging Flow Cytometry

Mitotic Golgi translocation of ERK1c is mediated by PI4KIIIβ/14-3-3γ shuttling complex

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