@phdthesis{daCruzGueerisoli2021,
  author    = {da Cruz G{\"u}erisoli, Irene Maria},
  title     = {Investigating the murine meiotic telomere complex TERB1-TERB2-MAJIN: spatial organization and evolutionary history},
  doi       = {10.25972/OPUS-21056},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-210562},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2021},
  abstract  = {Einess der faszinierenden Merkmale der meiotischen Prophase I sind die hochkonservierten kr{\"a}ftigen Bewegungen homologer Chromosomen. Diese Bewegungen sind entscheidend f{\"u}r den Erfolg von Schl{\"u}sselereignissen wie die Ausrichtung, Paarung und Rekombination der homologen Chromosomen. Mehrere bisher untersuchte Organismen, darunter S{\"a}ugetiere, W{\"u}rmer, Hefen und Pflanzen, erreichen diese Bewegungen, indem sie die Chromosomenenden an spezialisierten Stellen in der Kernh{\"u}lle verankern. Diese Verankerung erfordert Telomer-Adapterproteine, die bisher in der Spalthefe und der Maus identifiziert wurden. Die meiosespezifischen Telomer-Adapterproteine der Maus, TERB1, TERB2 und MAJIN, sind an der Verankerung des ubiquit{\"a}ren Telomer-Shelterin-protein an den LINC-Komplex beteiligt, mit einem analogen Mechanismus, wie er die Spalthefe beschrieben wird. Obgleich die meiose-spezifischen TelomerAdapterproteine eine wesentliche Rolle spielen, ist der genaue Mechanismus der Verankerung der Telomere an die Kernh{\"u}lle sowie ihre evolution{\"a}re Geschichte bisher noch wenig verstanden. Das Hauptziel dieser Arbeit ist daher die Untersuchung der Organisation des meiosespezifischen TelomerAdapterkomplexes TERB1-TERB2-MAJIN der Maus und dessen Evolutionsgeschichte. Im ersten Teil dieser Arbeit wurde die Organisation des TERB1-TERB2-MAJIN Komplexes mittels hochaufl{\"o}sender Mikroskopie (SIM), an Mausspermatozyten untersucht, sowie die Lokalisation in Bezug auf TRF1 des Telomer-ShelterinKomplexes und die telomerische DNA analysiert. In den Stadien Zygot{\"a}n und Pachyt{\"a}n zeigten die Fluoreszenzsignale eine starke {\"U}berlappung der Verteilung der meiotischen Telomer-Komplex-Proteine, wobei die Organisation von TERB2 an den Chromosomenenden heterogener war als die von TERB1 und MAJIN. Außerdem konnte die TRF1-Lokalisation an den Enden der Lateralelemente (LEs) mit einer griffartigen Anordnung um die TERB1- und MAJIN-Signale im Zygot{\"a}n- und Pachyt{\"a}n-Stadium gezeigt werden. Interessanterweise erwies sich die telomerische DNA als lateral verteilt und teilweise {\"u}berlappend mit der zentralen Verteilung der meiotischen Telomer-Komplex-Proteine an den Enden der LEs. Die Kombination dieser Ergebnisse erlaubte die Beschreibung eines alternativen Modells der Verankerung der Telomer an die Kernh{\"u}lle w{\"a}hrend der meiotischen Prophase I. Der zweite Teil dieser Arbeit analysiert die Evolutionsgeschichte der Mausproteine von TERB1, TERB2 und MAJIN. Die fehlende {\"U}bereinstimmung zwischen den Meiose-spezifische Telomer-Adapteproteinen der Maus und der Spalthefe hat die Frage nach dem evolutionsbedingten Ursprung dieses spezifischen Komplexes aufgeworfen. Um vermeintliche Orthologen der Mausproteinevon TERB1, TERB2 und MAJIN {\"u}ber Metazoen hinweg zu identifizieren, wurden computergest{\"u}tzte Verfahren und phylogenetische Analysen durchgef{\"u}hrt. Dar{\"u}ber hinaus wurden Expressionsstudien implementiert, um ihre potenzielle Funktion w{\"a}hrend der Meiose zu testen. Die Analysen haben ergeben, dass der Meiose-spezifische Telomer-Komplex der Maus sehr alt ist, da er bereits in den Eumetazoen entstand, was auf einen einzigen Ursprung hindeutet. Das Fehlen jeglicher Homologen des meiosespezifischen Telomerkomplexes in Nematoden und die einigen wenigen in Arthropoden nachgewiesenen Kandidaten, deuten darauf hin, dass die Telomer-Adapterproteine in diesen Abstammungslinien verloren/ersetzt oder stark diversifiziert worden sind. Bemerkenswerterweise zeigten Proteindom{\"a}nen von TERB1, TERB2 und MAJIN, die an der Bildung des Komplexes sowie an der Interaktion mit dem Telomer-Shelterin-Protein und den LINC-Komplexen beteiligt sind, eine hohe Sequenz{\"a}hnlichkeit {\"u}ber alle Kladen hinweg. Abschließend lieferte die Genexpression im Nesseltier Hydra vulgaris den Beweis, dass der TERB1-TERB2-MAJIN-Komplex selektiv in der Keimbahn exprimiert wird, was auf die Konservierung meiotischer Funktionen {\"u}ber die gesamte Metazoen-Evolution hinweg hindeutet. Zusammenfassend bietet diese Arbeit bedeutende neue Erkenntnisse hinsichtlich des Meiose-spezifischen Telomer-Adapterkomplex, seines Mechanismus zur Verankerung der Telomer an die Kernh{\"u}lle und die Entschl{\"u}sselung seines Ursprungs in den Metazoen.},
  language  = {en}
}
@article{daCruzRodriguezCasuriagaSantinaqueetal.2016,
  author    = {da Cruz, Irene and Rodr{\´i}guez-Casuriaga, Rosana and Santi{\~n}aque, Frederico F. and Far{\´i}as, Joaquina and Curti, Gianni and Capoano, Carlos A. and Folle, Gustavo A. and Benavente, Ricardo and Sotelo-Silveira, Jos{\´e} Roberto and Geisinger, Adriana},
  title     = {Transcriptome analysis of highly purified mouse spermatogenic cell populations: gene expression signatures switch from meiotic-to postmeiotic-related processes at pachytene stage},
  series = {BMC Genomics},
  volume    = {17},
  journal   = {BMC Genomics},
  doi       = {10.1186/s12864-016-2618-1},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-164574},
  pages     = {294},
  year      = {2016},
  abstract  = {Background Spermatogenesis is a complex differentiation process that involves the successive and simultaneous execution of three different gene expression programs: mitotic proliferation of spermatogonia, meiosis, and spermiogenesis. Testicular cell heterogeneity has hindered its molecular analyses. Moreover, the characterization of short, poorly represented cell stages such as initial meiotic prophase ones (leptotene and zygotene) has remained elusive, despite their crucial importance for understanding the fundamentals of meiosis. Results We have developed a flow cytometry-based approach for obtaining highly pure stage-specific spermatogenic cell populations, including early meiotic prophase. Here we combined this methodology with next generation sequencing, which enabled the analysis of meiotic and postmeiotic gene expression signatures in mouse with unprecedented reliability. Interestingly, we found that a considerable number of genes involved in early as well as late meiotic processes are already on at early meiotic prophase, with a high proportion of them being expressed only for the short time lapse of lepto-zygotene stages. Besides, we observed a massive change in gene expression patterns during medium meiotic prophase (pachytene) when mostly genes related to spermiogenesis and sperm function are already turned on. This indicates that the transcriptional switch from meiosis to post-meiosis takes place very early, during meiotic prophase, thus disclosing a higher incidence of post-transcriptional regulation in spermatogenesis than previously reported. Moreover, we found that a good proportion of the differential gene expression in spermiogenesis corresponds to up-regulation of genes whose expression starts earlier, at pachytene stage; this includes transition protein-and protamine-coding genes, which have long been claimed to switch on during spermiogenesis. In addition, our results afford new insights concerning X chromosome meiotic inactivation and reactivation. Conclusions This work provides for the first time an overview of the time course for the massive onset and turning off of the meiotic and spermiogenic genetic programs. Importantly, our data represent a highly reliable information set about gene expression in pure testicular cell populations including early meiotic prophase, for further data mining towards the elucidation of the molecular bases of male reproduction in mammals.},
  language  = {en}
}
@article{DedukhDaCruzKneitzetal.2022,
  author    = {Dedukh, Dmitrij and Da Cruz, Irene and Kneitz, Susanne and Marta, Anatolie and Ormanns, Jenny and Tichop{\´a}d, Tom{\´a}š and Lu, Yuan and Alsheimer, Manfred and Janko, Karel and Schartl, Manfred},
  title     = {Achiasmatic meiosis in the unisexual Amazon molly, Poecilia formosa},
  series = {Chromosome Research},
  volume    = {30},
  journal   = {Chromosome Research},
  number    = {4},
  doi       = {10.1007/s10577-022-09708-2},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-325128},
  pages     = {443-457},
  year      = {2022},
  abstract  = {Unisexual reproduction, which generates clonal offspring, is an alternative strategy to sexual breeding and occurs even in vertebrates. A wide range of non-sexual reproductive modes have been described, and one of the least understood questions is how such pathways emerged and how they mechanistically proceed. The Amazon molly, Poecilia formosa, needs sperm from males of related species to trigger the parthenogenetic development of diploid eggs. However, the mechanism, of how the unreduced female gametes are produced, remains unclear. Cytological analyses revealed that the chromosomes of primary oocytes initiate pachytene but do not proceed to bivalent formation and meiotic crossovers. Comparing ovary transcriptomes of P. formosa and its sexual parental species revealed expression levels of meiosis-specific genes deviating from P. mexicana but not from P. latipinna. Furthermore, several meiosis genes show biased expression towards one of the two alleles from the parental genomes. We infer from our data that in the Amazon molly diploid oocytes are generated by apomixis due to a failure in the synapsis of homologous chromosomes. The fact that this failure is not reflected in the differential expression of known meiosis genes suggests the underlying molecular mechanism may be dysregulation on the protein level or misexpression of a so far unknown meiosis gene, and/or hybrid dysgenesis because of compromised interaction of proteins from diverged genomes.},
  language  = {en}
}
@article{DunceMilburnGurusaranetal.2018,
  author    = {Dunce, James M. and Milburn, Amy E. and Gurusaran, Manickam and da Cruz, Irene and Sen, Lee T. and Benavente, Ricardo and Davies, Owen R.},
  title     = {Structural basis of meiotic telomere attachment to the nuclear envelope by MAJIN-TERB2-TERB1},
  series = {Nature Communications},
  volume    = {9},
  journal   = {Nature Communications},
  doi       = {10.1038/s41467-018-07794-7},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-226416},
  year      = {2018},
  abstract  = {Meiotic chromosomes undergo rapid prophase movements, which are thought to facilitate the formation of inter-homologue recombination intermediates that underlie synapsis, crossing over and segregation. The meiotic telomere complex (MAJIN, TERB1, TERB2) tethers telomere ends to the nuclear envelope and transmits cytoskeletal forces via the LINC complex to drive these rapid movements. Here, we report the molecular architecture of the meiotic telomere complex through the crystal structure of MAJIN-TERB2, together with light and X-ray scattering studies of wider complexes. The MAJIN-TERB2 2:2 hetero-tetramer binds strongly to DNA and is tethered through long flexible linkers to the inner nuclear membrane and two TRF1-binding 1:1 TERB2-TERB1 complexes. Our complementary structured illumination microscopy studies and biochemical findings reveal a telomere attachment mechanism in which MAJIN-TERB2-TERB1 recruits telomere-bound TRF1, which is then displaced during pachytene, allowing MAJIN-TERB2-TERB1 to bind telomeric DNA and form a mature attachment plate.},
  language  = {en}
}