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In eukaryotic cells, strategies have evolved to prevent conflicts between replication and transcription. Giardia lamblia, a binucleated protozoan, alternates between tetraploid and octaploid genomes during its cell cycle. Using single-molecule techniques like DNA combing and nanopore-based sequencing, we investigated the spatio-temporal organization of DNA replication, replication fork progression and potential head-on replication-transcription collisions in Giardia trophozoites. Our findings indicate that Giardia chromosomes are replicated from only a few active origins, which are widely spaced and exhibit faster replication rates compared to those in other protozoan parasites. Immunofluorescence assays revealed that \u223c20% of trophozoites show asynchronous replication between nuclei. Forksense and gene ontology analyses disclosed that genes in regions with potential head-on collisions are linked to chromatin dynamics, cell cycle regulation and DNA replication\/repair pathways, possibly explaining the observed asynchronous replication in part of the population. This study offers the first comprehensive view of replication dynamics in Giardia, which is the pathogen that causes giardiasis, a diarrheal disease impacting millions worldwide.<\/jats:p>","DOI":"10.1242\/jcs.260828","type":"journal-article","created":{"date-parts":[[2023,5,9]],"date-time":"2023-05-09T08:00:09Z","timestamp":1683619209000},"source":"Crossref","is-referenced-by-count":7,"title":["Clues on the dynamics of DNA replication in <i>Giardia lamblia<\/i>"],"prefix":"10.1242","volume":"136","author":[{"given":"Marcelo S.","family":"da Silva","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Ciclo celular, Instituto Butantan 1 , S\u00e3o Paulo, SP 05503-900 , Brazil"},{"name":"Centro de Toxinas, Resposta Imune e Sinaliza\u00e7\u00e3o Celular (CeTICS), Instituto Butantan 2 , S\u00e3o Paulo, SP 05503-900 , Brazil"},{"name":"Instituto de Qu\u00edmica, Universidade de S\u00e3o Paulo (USP) 3 Departamento de Bioqu\u00edmica , , S\u00e3o Paulo, SP 05508-900 , Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcela O.","family":"Vitarelli","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Ciclo celular, Instituto Butantan 1 , S\u00e3o Paulo, SP 05503-900 , Brazil"},{"name":"Centro de Toxinas, Resposta Imune e Sinaliza\u00e7\u00e3o Celular (CeTICS), Instituto Butantan 2 , S\u00e3o Paulo, SP 05503-900 , Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vincent Louis","family":"Viala","sequence":"additional","affiliation":[{"name":"Centro de Toxinas, Resposta Imune e Sinaliza\u00e7\u00e3o Celular (CeTICS), Instituto Butantan 2 , S\u00e3o Paulo, SP 05503-900 , Brazil"},{"name":"Laborat\u00f3rio de Bioqu\u00edmica, Instituto Butantan 4 , S\u00e3o Paulo, SP 05503-900 , Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8497-706X","authenticated-orcid":false,"given":"Katherine","family":"Tsantarlis","sequence":"additional","affiliation":[{"name":"Universidade Federal de S\u00e3o Paulo 5 Departamento de Microbiologia, Imunologia e Parasitologia , , S\u00e3o Paulo, SP 04023-062 , Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4662-6469","authenticated-orcid":false,"given":"David","family":"da Silva Pires","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Ciclo celular, Instituto Butantan 1 , S\u00e3o Paulo, SP 05503-900 , Brazil"},{"name":"Centro de Toxinas, Resposta Imune e Sinaliza\u00e7\u00e3o Celular (CeTICS), Instituto Butantan 2 , S\u00e3o Paulo, SP 05503-900 , Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thiago A.","family":"Franco","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Ciclo celular, Instituto Butantan 1 , S\u00e3o Paulo, SP 05503-900 , Brazil"},{"name":"Centro de Toxinas, Resposta Imune e Sinaliza\u00e7\u00e3o Celular (CeTICS), Instituto Butantan 2 , S\u00e3o Paulo, SP 05503-900 , Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Inacio L. 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