title: TMF/ARA160 Governs the Dynamic Spatial Orientation of the Golgi Apparatus during Sperm Development.
abstract: TMF/ARA160 is known to be a TATA element Modulatory Factor (TMF). It was initially identified as a DNA-binding factor and a coactivator of the Androgen receptor. It was also characterized as a Golgi-associated protein, which is essential for acrosome formation during functional sperm development. However, the molecular roles of TMF in this intricate process have not been revealed. Here, we show that during spermiogenesis, TMF undergoes a dynamic change of localization throughout the Golgi apparatus. Specifically, TMF translocates from the cis-Golgi to the trans-Golgi network and to the emerging vesicles surface, as the round spermatids develop. Notably, lack of TMF led to an abnormal spatial orientation of the Golgi and to the deviation of the trans-Golgi surface away from the nucleus of the developing round spermatids. Concomitantly, pro-acrosomal vesicles derived from the TMF-/- Golgi lacked targeting properties and did not tether to the spermatid nuclear membrane thereby failing to form the acrosome anchoring scaffold, the acroplaxome, around the cell-nucleus. Absence of TMF also perturbed the positioning of microtubules, which normally lie in proximity to the Golgi and are important for maintaining Golgi spatial orientation and dynamics and for chromatoid body formation, which is impaired in TMF-/- spermatids. In-silico evaluation combined with molecular and electron microscopic analyses revealed the presence of a microtubule interacting domain (MIT) in TMF, and confirmed the association of TMF with microtubules in spermatogenic cells. Furthermore, the MIT domain in TMF, along with microtubules integrity, are required for stable association of TMF with the Golgi apparatus. Collectively, we show here for the first time that a Golgi and microtubules associated protein is crucial for maintaining proper Golgi orientation during a cell developmental process.
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel.
authors.forename: Yoav
authors.initials: Y
authors.lastname: Elkis
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel.
authors.forename: Shai
authors.initials: S
authors.lastname: Bel
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel.
authors.forename: Roni
authors.initials: R
authors.lastname: Rahimi
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel.
authors.forename: Tali
authors.initials: T
authors.lastname: Lerer-Goldstein
authors.affiliation: Electron Microscopy Unit, Weizmann Institute of Science, Rehovot, 7610001, Israel.
authors.forename: Smadar
authors.initials: S
authors.lastname: Levin-Zaidman
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Babushkin
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel.
authors.forename: Sally
authors.initials: S
authors.lastname: Shpungin
authors.affiliation: The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, 52900, Israel.
authors.forename: Uri
authors.initials: U
authors.lastname: Nir
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D004268:DNA-Binding Proteins; D017353:Gene Deletion; D006056:Golgi Apparatus / Q000378:metabolism*; D000077614:Golgi Matrix Proteins; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D041681:NIH 3T3 Cells; D017434:Protein Structure, Tertiary; D020539:Sequence Analysis, Protein; D013087:Spermatids / Q000378:metabolism / Q000648:ultrastructure; D013091:Spermatogenesis; D014157:Transcription Factors; D014404:Tubulin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000502:physiology*; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D000077614:Golgi Matrix Proteins; C554202:Tmf1 protein, mouse; D014157:Transcription Factors; D014404:Tubulin; D033921:Vesicular Transport Proteins; D044767:Ubiquitin-Protein Ligases