title: The large noncoding hsrω-n transcripts are essential for thermotolerance and remobilization of hnRNPs, HP1 and RNA polymerase II during recovery from heat shock in Drosophila.
abstract: The hs-GAL4(t)-driven expression of the hsrω-RNAi transgene or EP93D allele of the noncoding hsrω resulted in global down- or upregulation, respectively, of the large hsrω-n transcripts following heat shock. Subsequent to temperature shock, hsrω-null or those expressing hsrω-RNAi or the EP93D allele displayed delayed lethality of most embryos, first or third instar larvae. Three-day-old hsrω-null flies mostly died immediately or within a day after heat shock. Heat-shock-induced RNAi or EP expression in flies caused only a marginal lethality but severely affected oogenesis. EP allele or hsrω-RNAi expression after heat shock did not affect heat shock puffs and Hsp70 synthesis. Both down- and upregulation of hsrω-n transcripts suppressed reappearance of the hsrω-n transcript-dependent nucleoplasmic omega speckles during recovery from heat shock. Hrp36, heterochromatin protein 1, and active RNA pol II in unstressed or heat-shocked wild-type or hsrω-null larvae or those expressing the hs-GAL4(t)-driven hsrω-RNAi or the EP93D allele were comparably distributed on polytene chromosomes. Redistribution of these proteins to pre-stress locations after a 1- or 2-h recovery was severely compromised in glands with down- or upregulated levels of hsrω-n transcripts after heat shock. The hsrω-null unstressed cells always lacked omega speckles and little Hrp36 moved to any chromosome region following heat shock, and its relocation to chromosome regions during recovery was also incomplete. This present study reveals for the first time that the spatial restoration of key regulatory factors like hnRNPs, HP1, or RNA pol II to their pre-stress nuclear targets in cells recovering from thermal stress is dependent upon critical level of the large hsrω-n noncoding RNA. In the absence of their relocation to pre-stress chromosome sites, normal developmental gene activity fails to be restored, which finally results in delayed organismal death.
authors.affiliation: Cytogenetics Laboratory, Department of Zoology, Banaras Hindu University, Varanasi 221005, India. lakhotia@bhu.ac.in
authors.forename: Subhash C
authors.initials: SC
authors.lastname: Lakhotia
authors.forename: Moushami
authors.initials: M
authors.lastname: Mallik
authors.forename: Anand K
authors.initials: AK
authors.lastname: Singh
authors.forename: Mukulika
authors.initials: M
authors.lastname: Ray
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics* / Q000502:physiology; D000818:Animals; D030801:Animals, Genetically Modified; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D004330:Drosophila / Q000196:embryology / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D029721:Drosophila Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D005838:Genotype; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics*; D034441:Heterogeneous-Nuclear Ribonucleoproteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D009687:Nuclear Proteins; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D022661:RNA, Untranslated / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013696:Temperature
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D029721:Drosophila Proteins; D006360:Heat-Shock Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; C402657:Hrb87F protein, Drosophila; D009687:Nuclear Proteins; D022661:RNA, Untranslated; C452193:heterochromatin protein 1, Drosophila; D012319:RNA Polymerase II