Dephosphorylation of YB-1 is Required for Nuclear Localisation During G2 Phase of the Cell Cycle

Mehta, Sunali; McKinney, Cushla; Algie, Michael; Verma, Chandra; Kannan, Srinivasaraghavan; Harfoot, Rhodri; Bartolec, Tara K.; Bhatia, Puja; Fisher, Alistair J.; Gould, Maree; Parker, Kim H.; Cesare, Anthony J.; Cunliffe, Heather E.; Cohen, Scott B.; Kleffmann, Torsten; Braithwaite, Antony W.; Woolley, Adele G.

doi:10.3390/cancers12020315

Cited by 19 publications

(18 citation statements)

References 58 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This suggests that YB-1 is required for formation of the cleavage furrow. To test this, we first used confocal microscopy to determine whether YB-1 is localized at or near the cleavage furrow, using a validated, highly specific antibody to the N-terminus of YB-1 [ 5 , 41 , 42 ] (and Figure S1a,b ). Results ( Figure 2 c) show that as expected AURKB (a marker of CPC) is enriched at the spindle mid-zone.…”

Section: Resultsmentioning

confidence: 99%

“…YB-1 function is thought to be regulated through phosphorylation [ 42 ]. Inhibiting YB-1 phosphorylation on serine 102 with a mutation to alanine (S102A) prevented cell proliferation [ 48 , 49 ] and transactivation of various genes including EGFR [ 50 ] and PIK3CA [ 51 ] and NOTCH4 [ 52 ].…”

Section: Resultsmentioning

confidence: 99%

“…YB-1 was immunopurified as described previously by Cohen et al [ 42 ]. Lysates were prepared at the equivalent of 1 × 10 4 cells per µL for whole cells or cytoplasmic lysates and 5 × 10 4 cells per µL for nuclear lysates in a low detergent RIPA buffer (50 mM Tris-HCL, 150 mM NaCl, 0.5% NP40, 0.1% deoxycholate).…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Critical Role for Cold Shock Protein YB-1 in Cytokinesis

Mehta

Algie

Al-Jabry

et al. 2020

Cancers

Self Cite

View full text Add to dashboard Cite

High levels of the cold shock protein Y-box-binding protein-1, YB-1, are tightly correlated with increased cell proliferation and progression. However, the precise mechanism by which YB-1 regulates proliferation is unknown. Here, we found that YB-1 depletion in several cancer cell lines and in immortalized fibroblasts resulted in cytokinesis failure and consequent multinucleation. Rescue experiments indicated that YB-1 was required for completion of cytokinesis. Using confocal imaging we found that YB-1 was essential for orchestrating the spatio-temporal distribution of the microtubules, β-actin and the chromosome passenger complex (CPC) to define the cleavage plane. We show that phosphorylation at six serine residues was essential for cytokinesis, of which novel sites were identified using mass spectrometry. Using atomistic modelling we show how phosphorylation at multiple sites alters YB-1 conformation, allowing it to interact with protein partners. Our results establish phosphorylated YB-1 as a critical regulator of cytokinesis, defining precisely how YB-1 regulates cell division.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Critical Role for Cold Shock Protein YB-1 in Cytokinesis

Mehta

Algie

Al-Jabry

et al. 2020

Cancers

Self Cite

View full text Add to dashboard Cite

show abstract

“…Analyses by NMR and isothermal titration calorimetry (ITC) of DNA-heptamer binding to an extended YBX1 CSD reveals DNA-sequence preferences of YBX1, a binding mode resembling bacterial CSPs and the structural basis of the observed attenuated target DNA binding by S102 phosphorylation [ 9 ]. Conversely, dephosphorylation of S102 and other serine residues was proposed to unmask the nuclear localization signal of YBX1 and facilitate nuclear entry at specific stages during the cell cycle [ 141 ]. Protein kinase AKT-mediated phosphorylation of YBX1 also regulates its binding to the capped 5′-end of mRNA [ 142 ].…”

Section: Cold-shock Domain-binding To Nucleic Acidsmentioning

confidence: 99%

Cold-Shock Domains—Abundance, Structure, Properties, and Nucleic-Acid Binding

Heinemann

Roske

2021

Cancers

View full text Add to dashboard Cite

The cold-shock domain has a deceptively simple architecture but supports a complex biology. It is conserved from bacteria to man and has representatives in all kingdoms of life. Bacterial cold-shock proteins consist of a single cold-shock domain and some, but not all are induced by cold shock. Cold-shock domains in human proteins are often associated with natively unfolded protein segments and more rarely with other folded domains. Cold-shock proteins and domains share a five-stranded all-antiparallel β-barrel structure and a conserved surface that binds single-stranded nucleic acids, predominantly by stacking interactions between nucleobases and aromatic protein sidechains. This conserved binding mode explains the cold-shock domains’ ability to associate with both DNA and RNA strands and their limited sequence selectivity. The promiscuous DNA and RNA binding provides a rationale for the ability of cold-shock domain-containing proteins to function in transcription regulation and DNA-damage repair as well as in regulating splicing, translation, mRNA stability and RNA sequestration.

show abstract

“…p90 Ribosomal-S6 Kinase (RSK) phosphorylates YB-1 and is required for cytokinesis. YB-1 function is thought to be regulated through phosphorylation 38 . Inhibiting YB-1 phosphorylation on serine 102 with a mutation to alanine (S102A) prevented cell proliferation 39 and transactivation of various genes including EGFR 40 and PIK3CA 41 .…”

Section: Yb-1 Depletion Causes Cytokinesis Failure In Zebrafish Embrymentioning

confidence: 99%

Critical role for cold shock protein YB-1 in cytokinesis

Mehta

Algie

Al-Jabri

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Y-box-binding protein-1, YB-1, is essential for cell division, but it is not clear how it functions. Using live imaging and confocal microscopy we show that YB-1 functions only in the last step of division, specifically being required to initiate cytokinesis. ABSTRACTHigh levels of the cold shock protein Y-box-binding protein-1, YB-1, are tightly correlated with increased cell proliferation and cancer progression. However, the precise mechanism by which YB-1 regulates proliferation is unknown. Here, we found that YB-1 depletion in several cell lines resulted in cytokinesis failure, multinucleation and an increase in G1 transit time. Rescue experiments indicated that YB-1 was required for completion of cytokinesis. Using confocal imaging of cells undergoing cytokinesis both in vitro and in zebrafish embryos, we found that YB-1 was critical for microtubule organization during cytokinesis. Using mass spectrometry we identified multiple novel phosphorylation sites on YB-1. We show that phosphorylation of YB-1 at multiple serine residues was essential for its function during cytokinesis. Using atomistic modelling we show how multiple phosphorylations alter YB-1 conformation, allowing it to interact with protein partners. Our results establish phosphorylated YB-1 as a critical regulator of cytokinesis, defining for the first time precisely how YB-1 regulates cell division.Word Count 157 / 160 words

show abstract

Dephosphorylation of YB-1 is Required for Nuclear Localisation During G2 Phase of the Cell Cycle

Cited by 19 publications

References 58 publications

Critical Role for Cold Shock Protein YB-1 in Cytokinesis

Critical Role for Cold Shock Protein YB-1 in Cytokinesis

Cold-Shock Domains—Abundance, Structure, Properties, and Nucleic-Acid Binding

Critical role for cold shock protein YB-1 in cytokinesis

Contact Info

Product

Resources

About