Track topics on Twitter Track topics that are important to you
XMAP215/Stu2/Alp14 accelerates tubulin polymerization while processively tracking microtubule plus-ends via Tumor Overexpressed Gene (TOG) domain arrays. It remains poorly understood how these functions arise from tubulin recruitment, mediated by the distinct TOG1 and TOG2 domains, or the assembly of these arrays into large square complexes. Here, we describe a relationship between microtubule plus-end tracking and polymerase functions revealing their distinct origin within TOG arrays. We study Alp14 mutants designed based on structural models, with defects in either tubulin recruitment or self-organization. Using in vivo live imaging in fission yeast and in vitro microtubule dynamics assays, we show that tubulins recruited by TOG1 and TOG2 serve concerted, yet distinct, roles in microtubule plus-end tracking and polymerase functions. TOG1 is critical for processive plus-end tracking while TOG2 is critical for accelerating tubulin polymerization. Inactivating interfaces that stabilize square complexes lead to defects in both processive microtubule plus-end tracking and polymerase. Our studies suggest that a dynamic cycle between square and unfurled TOG array states gives rise to processive polymerase activity at microtubule plus-ends. Movie S1 Movie S1 WT-Alp14-NG compared to TOG1M-NG and TOG2M-NG in vitro experiments Left, two representative movies of WT-Alp14-mNG tracking dynamic MTs. Middle, two representative movies of Alp14 TOG1M-NG tracking dynamic MTs. Left, two representative movies of Alp14 TOG2M-NG tracking dynamic MTs. Star marks signal at the MT plus end. Movie S2 Movie S2 Single molecule experiments revealing the residence of wt-Alp14-SNAP-Atto-488 compared to TOG1M-SNAP-Atto-488 single TOG2M-SNAP at MT plus ends. Left, Representative movies of WT Alp14-SNAP-Atto-488 tracking dynamic MTs.. Middle, representative movies of Alp14 TOG1M-SNAP-Atto-488 tracking dynamic MTs. Right, representative movies of Alp14 TOG2M-SNAP-Atto-488 tracking dynamic MTs. Star marks signal at the MT plus end. Movie S3 Movie S3 INT1-NG compared to INT2-NG and INT1+2-NG in vitro experiments Left, two representative movies of INT1-mNG tracking dynamic MTs. Star marks signal at the MT plus end. Middle, two representative movies of INT2-mNG tracking dynamic MTs. Star marks signal at the MT plus end. Left, two representative movies of INT1+2-mNG tracking dynamic MTs. Star marks signal at the MT plus end. Movie S4 Movie S4 Single molecule experiments revealing the residence of INT1-SNAP-Atto-488, INT2-SNAP-Atto-488 single, INT1+2-SNAP-Atto-488 at MT plus ends. Left, Representative movies of INT1-SNAP-Atto-488 tracking dynamic MTs.. Middle, representative movies of INT2-SNAP-Atto-488 tracking dynamic MTs. Right, representative movies of INT1+2-SNAP-Atto-488 tracking dynamic MTs. Star marks signal at the MT plus end.
This article was published in the following journal.
Name: Molecular biology of the cell
End-binding proteins (EBs), referred to as the core components of the microtubule plus-end tracking protein network, interact with the C-terminus of the Adenomatous Polyposis Coli (APC) tumor suppress...
The DNA polymerase δ catalytic subunit (PolD1) is a highly conserved protein with established functions in both the nucleus and the cytoplasm: whereas PolD1 participates in the replication and repair...
Dynamic control of the actin and microtubule cytoskeletons underlie nearly every critical process during neural development, and requires multiple dimensions of regulation. Formins are a family of fif...
Microtubule-severing enzymes, which can remove tubulin dimers from microtubule lattices, participate in cytoskeletal remodeling in various contexts. A recent study showed that partially damaged microt...
Cytoplasmic dynein-1 (hereafter dynein) is an essential cellular motor that drives the movement of diverse cargos along the microtubule cytoskeleton, including organelles, vesicles and RNAs. A long-st...
This phase I trial studies the side effects and best dose of microtubule-targeted agent BAL101553 when given together with radiation therapy in treating patients with newly diagnosed gliob...
This pilot study examines concurrent and predictive relationships between eye tracking and clinical outcomes during a 16-week behavioral intervention (PRT) for children with ASD. Eye track...
Clarify the relation of microtubule-associated protein 2 and cell migration
This study will assess the effects of exercise and non-concussive bodily contact on eye-tracking scores collected by the EYE-SYNC eye-tracking device.
Numerous studies have demonstrated that excess perivisceral adipose tissue is associated with metabolic diseases such as insulin resistance. In skeletal muscle, insulin resistance has bee...
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. The enzyme functions in the nucleolar structure and transcribes DNA into RNA. It has different requirements for cations and salts than RNA polymerase II and III and is not inhibited by alpha-amanitin. EC 188.8.131.52.
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure and transcribes DNA into RNA. It has different requirements for cations and salt than RNA polymerase I and is strongly inhibited by alpha-amanitin. EC 184.108.40.206.
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure where it transcribes DNA into RNA. It has specific requirements for cations and salt and has shown an intermediate sensitivity to alpha-amanitin in comparison to RNA polymerase I and II. EC 220.127.116.11.
A DNA-dependent DNA polymerase characterized in E. coli and other lower organisms but may be present in higher organisms. Use also for a more complex form of DNA polymerase III designated as DNA polymerase III* or pol III* which is 15 times more active biologically than DNA polymerase I in the synthesis of DNA. This polymerase has both 3'-5' and 5'-3' exonuclease activities, is inhibited by sulfhydryl reagents, and has the same template-primer dependence as pol II. EC 18.104.22.168.
A DNA-directed DNA polymerase that functions in the replication of MITOCHONDRIAL DNA. Mutations in the gene that encodes this enzyme (POLG) are associated with some forms of OPHTHALMOPLEGIA, CHRONIC EXTERNAL PROGRESSIVE.
Polymerase Chain Reaction (PCR)
PCR (Polymerase Chain Reaction) uses the ability of DNA polymerase (enzymes that create DNA molecules by assembling nucleotides, the building blocks of DNA. These enzymes are essential to DNA replication and usually work in pairs to create two ident...
Bioinformatics is the application of computer software and hardware to the management of biological data to create useful information. Computers are used to gather, store, analyze and integrate biological and genetic information which can then be applied...
Immunoassay - ELISA
Immunoassays are quick and accurate tests to detect specific molecules. Immunoassays rely on an antibody to bind to the specific structure of a molecule. Antibodies are proteins generated by animals in response to the invasion of a foreign molecule (anti...