A phosphorylation-controlled switch confers cell cycle-dependent protein relocalization.
Tools for acute manipulation of protein localization enable elucidation of spatiotemporally defined functions, but their reliance on exogenous triggers can interfere with cell physiology. This limitation is particularly apparent for studying mitosis, whose highly choreographed events are sensitive to perturbations. Here we exploit the serendipitous discovery of a phosphorylation-controlled, cell cycle-dependent localization change of the adaptor protein PLEKHA5 to develop a system for mitosis-specific protein recruitment to the plasma membrane that requires no exogenous stimulus. Mitosis-enabled Anchor-away/Recruiter System (MARS) comprises an engineered, 15-kDa module derived from PLEKHA5 capable of recruiting functional protein cargoes to the plasma membrane during mitosis, either through direct fusion or via GFP-GFP nanobody interaction. Applications of MARS include both knock sideways to rapidly extract proteins from their native localizations during mitosis and conditional recruitment of lipid-metabolizing enzymes for mitosis-selective editing of plasma membrane lipid content, without the need for exogenous triggers or perturbative synchronization methods.
Pubmed ID: 38895347 RIS Download
Research resources used in this publication
None foundAdditional research tools detected in this publication
- Addgene (RRID:SCR_002037)
- Fiji (RRID:SCR_002285)
- Novus Biologicals (RRID:SCR_004286)
- Promega (RRID:SCR_006724)
- Thermo Fisher Scientific (RRID:SCR_008452)
- Santa Cruz Biotechnology (RRID:SCR_008987)
- Sigma-Aldrich (RRID:SCR_008988)
- Program to Reduce Incontinence by Diet and Exercise (RRID:SCR_009018)
- CoMet (RRID:SCR_011925)
- Thermo Xcalibur (RRID:SCR_014593)
Antibodies used in this publication
- Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 (RRID:AB_141607)
- mCherry Antibody (RRID:AB_11034849)
- GFP (B-2) (RRID:AB_627695)
- Histone H3 (D1H2) XP Rabbit mAb (RRID:AB_10544537)
- Monoclonal Anti-alpha-Tubulin antibody produced in mouse (RRID:AB_477579)
- Peroxidase-AffiniPure Goat Anti-Rabbit IgG (H+L) (min X Hu,Ms,Rat Sr Prot) (RRID:AB_2307391)
- Peroxidase-AffiniPure Goat Anti-Mouse IgG (H+L) (min X Hu,Bov,Hrs,Rb,Sw Sr Prot) (RRID:AB_2307392)
- EEA1 Polyclonal Antibody (RRID:AB_2096819)
- Mouse Anti-Human CD44 Monoclonal Antibody, Unconjugated, Clone 156-3C11 (RRID:AB_2076465)
- MOUSE ANTI V5-TAG (RRID:AB_567249)
- Anti-Actin, mouse monoclonal antibody (Clone C4) (RRID:AB_2335127)
- Phospho-Threonine/Tyrosine Antibody (RRID:AB_330301)
- Anti-EGFR (RABBIT) Antibody - 100-401-149 (RRID:AB_2293276)
Associated grants
- Agency: NIGMS NIH HHS, United States
Id: R01 GM143367 - Agency: NIGMS NIH HHS, United States
Id: R35 GM141159 - Agency: NIGMS NIH HHS, United States
Id: T32 GM138826
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This is a list of tools and resources that we have found mentioned in this publication.
Addgene (tool)
RRID:SCR_002037
Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
View all literature mentionsFiji (tool)
RRID:SCR_002285
Software package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.
View all literature mentionsNovus Biologicals (tool)
RRID:SCR_004286
Commercial antibody vendor which supplies antibodies and other products to life science researchers.
View all literature mentionsThermo Fisher Scientific (tool)
RRID:SCR_008452
Commercial vendor and service provider of laboratory reagents and antibodies. Supplier of scientific instrumentation, reagents and consumables, and software services.
View all literature mentionsSigma-Aldrich (tool)
RRID:SCR_008988
American chemical, life science and biotechnology company owned by Merck KGaA. Merger of Sigma Chemical Company and Aldrich Chemical Company. Provides organic and inorganic chemicals, building blocks, reagents, advanced materials and stable isotopes for chemical synthesis, medicinal chemistry and materials science, antibiotics, buffers, carbohydrates, enzymes, forensic tools, hematology and histology, nucleotides, proteins, peptides, amino acids and their derivatives.
View all literature mentionsProgram to Reduce Incontinence by Diet and Exercise (tool)
RRID:SCR_009018
Randomized controlled trial being conducted at two clinical centers in the United States to learn more about the effects of weight loss on urinary incontinence. About 330 overweight women aged 30 or older will participate and will be followed for 18 months. Efficacy of weight reduction as a treatment for urinary incontinence will be examined at 6 months following the intensive weight control program, and the sustained impact of the intervention will be examined at 18 months. To increase the maintenance of weight reduction and facilitate evaluation of the enduring impact of weight loss on urinary incontinence, they propose to study a motivation-based weight maintenance program. At the end of the intensive weight control program, women randomized to the weight loss program will be randomized to either a 12-month skill-based maintenance intervention or to a motivation-based maintenance intervention. The maintenance interventions maximize the potential for sustained weight loss and will allow them to determine if long-term weight reduction will produce continued improvement in urinary incontinence.
View all literature mentionsCoMet (tool)
RRID:SCR_011925
A web-server for fast comparative functional profiling of metagenomes.
View all literature mentionsThermo Xcalibur (tool)
RRID:SCR_014593
A software which acquires and processes data sets, primarily through the Xcalibur system.
View all literature mentionsDonkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 (antibody)
RRID:AB_141607
This polyclonal secondary targets IgG (H+L)
View all literature mentionsmCherry Antibody (antibody)
RRID:AB_11034849
This monoclonal targets mCherry
View all literature mentionsHistone H3 (D1H2) XP Rabbit mAb (antibody)
RRID:AB_10544537
This monoclonal targets Histone H3 (D1H2) XP Rabbit mAb
View all literature mentionsMonoclonal Anti-alpha-Tubulin antibody produced in mouse (antibody)
RRID:AB_477579
This monoclonal targets α-tubulin
View all literature mentionsPeroxidase-AffiniPure Goat Anti-Rabbit IgG (H+L) (min X Hu,Ms,Rat Sr Prot) (antibody)
RRID:AB_2307391
This polyclonal targets Rabbit IgG (H+L)
View all literature mentionsPeroxidase-AffiniPure Goat Anti-Mouse IgG (H+L) (min X Hu,Bov,Hrs,Rb,Sw Sr Prot) (antibody)
RRID:AB_2307392
This polyclonal targets Mouse IgG (H+L)
View all literature mentionsEEA1 Polyclonal Antibody (antibody)
RRID:AB_2096819
This unknown targets EEA1
View all literature mentionsMouse Anti-Human CD44 Monoclonal Antibody, Unconjugated, Clone 156-3C11 (antibody)
RRID:AB_2076465
This monoclonal targets Human CD44
View all literature mentionsMOUSE ANTI V5-TAG (antibody)
RRID:AB_567249
This monoclonal targets MOUSE ANTI V5-TAG
View all literature mentionsAnti-Actin, mouse monoclonal antibody (Clone C4) (antibody)
RRID:AB_2335127
This monoclonal targets Actin
View all literature mentionsPhospho-Threonine/Tyrosine Antibody (antibody)
RRID:AB_330301
This polyclonal targets Phospho-Threonine/Tyrosine
View all literature mentionsAnti-EGFR (RABBIT) Antibody - 100-401-149 (antibody)
RRID:AB_2293276
This unknown targets EGFR Antibody
View all literature mentions
