Aquesta tesi es centra en la caracterització funcional d'una proteïna de xoc de calor de baix pes molecular (Small Heat Shock Protein - sHSP) de classe I de surera pel que fa a la seva capacitat per protegir les cèl·lules de l'estrès i per estabilitzar les membranes. Les sHsps són proteïnes que s'expressen en condicions d'estrès cel·lular. Encara que certs aspectes funcionals de les sHsps són ben coneguts, el nostre treball aporta informacions noves sobre el paper de les diferents regions de la proteïna, especialment de la regió N-terminal.L'objectiu concret d'aquest treball és determinar la funció termoprotectora de QsHsp17.4-CI, una sHsp de classe I oobtinguda a partir de les cèl·lules de fel·lema d'alzina surera, en un model bacterià i analitzar la importància de les diferents regions de la proteïna en aquesta funció. Amb aquesta finalitat s'han dissenyat dues proteïnes parcials derivades de QsHsp17.4-CI: una a la que li falta la regió N-terminal (C105) i una altra amb pràcticament tot el domini -cristal·lí deleccionat (N61), i una tercera, derivada de QsHs10-CI, a la que li falta la meitat del domini -cristal·lí (Hsp10). També s'estudia la possible capacitat estabilitzadora de membranes i la capacitat de modificar l'expressió d'altres Hsps quan s'expressa de forma heteròloga.Els nostres resultats demostren que l'expressió de QsHsp17.4-CI protegeix a les cèl·lules d'E.coli de l'estrès tèrmic alhora que la regió N-terminal i la regió consens II del domini -cristal·lí són imprescindibles per aquesta funció de protecció. En relació a un possible paper en les membranes, els estudis de localització subcel·lular mostren que QsHsp17.4-CI colocalitza amb la fracció membranes i que la regió N-terminal de la proteïna és responsable d'aquesta colocalització. No s'ha pogut demostrar, però, que la localització amb la membrana estigui associada a un efecte protector d'aquesta: en cap cas la sobrexpressió de les proteïnes modifica la composició d'àcids grassos i només N61, que no té acció termoprotectora, altera l'estat fisico-químic de la membrana. En estudis d'expressió de novo en E.coli s'ha observat que, a diferència de les altres proteïnes heteròlogues, N61 activa l'expressió de la majoria de Hsps d'E.coli fent pensar en una possible relació entre l'estat físic de la membrana i l'activació de la resposta a l'estrès.En resum, en aquest treball hem provat la capacitat protectora de QsHsp17.4 i aportem noves dades sobre la importància de la regió N-terminal i la regió consens II del domini -cristal·lí en aquesta funció. Per altra banda, es suggereix que QsHsp17.4 podria interaccionar amb la membrana d'E.coli i que la regió N-terminal seria imprescindible per aquesta interacció. Finalment hem determinat que les proteïnes que provoquen variacions en l'estat de fluïdesa de la membrana poden activar la resposta al xoc de calor per part de la cèl·lula bacteriana. / This thesis is focused in the functional studies of a Small Heat Shock Protein (sHsp). sHsps are expressed under stress conditions. Although some functional aspects of these proteins are known, our work aport new data about the role of the different protein regions, especially the N-terminal region. The aim of this work is to demonstrate a thermotolerance effect of QsHsp17.4-CI in bacterial cells and to analyze the importance of the protein regions in this function. To achieve this objective two deletion mutants derived from QsHsp17.4-CI were designed: a protein lacking the N-terminal region (C105) and a protein where the entire -cristallin domain is missing (N61) and a third mutant, derived from QsHsp10-CI, that bears half of the -cristallin domain (Hsp10). To better understand the functional mechanism of sHsps we study the membrane stabilizing capacity of QsHsp17.4-CI as well as its capacity to modify other Hsps expression.Our results demonstrate that the expression of QsHsp17.4-CI protects E.coli cells from a heat shock and that the N-terminal region and the consensus region II of the -cristallin domain are necessary for this protective function. Related to a possible role in membranes, location studies suggest that QsHsp17.4-CI colocalizes with cell membrane fraction and that N-terminal region is important for this location. However, no relation between membrane localization and a protective effect has been demonstrated: Protein overexpression does not modify membrane fatty acid composition and only N61, which has no thermoprotection, changes membrane physical state. Studies of E.coli de novo synthesis show that, unlike the other recombinant proteins, the overexpression of N61 activates the expression of almost all E.coli Hsps suggesting a possible relation between membrane physical state and the activation of the heat shock response.As summary, in this work we have demonstrated the thermoprotective capacity of QsHsp17.4-CI and we contribute with new data about the importance of N-terminal region and consensus region II of -cristallin domain for this function. On the other hand, we suggest the possibility that QsHsp17.4-CI interacts with membrane and that N-terminal region is important for this interaction. Lastly, we have observed how changes in membranes fluidity state can activate heat shock response in bacterial cells.

There is evidence that athermal radiofrequency radiation can alter Heat Shock Protein (HSP) expression or protein phosphorylation, or alter MAP kinase signalling. Effects of long-term exposure in brain tissue due to repeated HSP perturbation (eg an inhibition of apoptosis) have been hypothesised (French et al, 2001). This study aimed to investigate the RNA expression profile (12,000 genes) and HSP family protein expression levels after either acute 1-hour or chronic 4-day intermittent exposures to simulated GSM radiation in a human primary fibroblast model. The results found minimal or no effects of GSM. Flasks were exposed to 900 MHz (217 Hz modulation) at 0.18 W/kg SAR within a Transverse Electromagnetic Mode chamber (TEM cell). Cultures rested for 2 hours before exposures. Affymetrix U95A microarray analysis of a single pilot set of experiments showed that about 40 genes were reported as upregulated &gt=2.5 fold in each condition. There was no evidence of altered expression of any MAPK-associated genes. Target genes reported in both conditions (CBFA2T1, ZNF148, ITGA1), and genes altered in one condition (CCS, PLEC1, BIRC5), and marginally altered HSP72 were selected for PCR analysis. No other members of the HSP family were altered. In three replicate experiments assayed by real-time PCR, six genes were either unchanged or showed randomly variable expression. However HSP72 RNA showed possible consistent slight upregulation of 1.37 +/- 0.21 in the chronic condition. Western immunoblots of HSP-60, -70, -72 and -V90 proteins showed no significant changes 5 hours after exposure. In preliminary studies using a serum starvation protocol, ERK-1 phosphorylation was unaltered after 5 or 30 minutes GSM (single experiments). When flasks were transiently cooled, ERK-1 phosphorylation was increased 20 minutes later, indicating a source of artefact in some protocols. An inflammatory challenge experiment with a low-dose of the cytokine IL-1???? found that acute GSM exposure post-challenge inhibited NF????B-mediated GRO???? induction by 1.5 fold (2 experiments). Preconditioning with mild heat induces transient inhibition of both NF????B signalling and apoptosis. Other studies indicate that EMF exposures similarly evoke cytoprotection. It is suggested that GSM evoked cytoprotective signalling in this inflammatory model.

[Truncated abstract] Breast cancer is the cause of significant suffering and death in our community. It is now estimated that the risk of developing breast cancer for an Australian woman before the age of 85 is 1 in 8, with this risk rising for unknown reasons. While mortality rates from breast cancer are falling due to increased awareness and early detection, few new treatments have been developed from an advanced understanding of the molecular basis of the disease. From decades of scientific research it is clear that estrogen (E2) has a large role to play in breast cancer. However, the basic mechanism behind E2 action in breast cancer remains unclear. E2 plays a fundamental role in breast cancer cell proliferation and is highly expressed in breast cancers, thus, it is important to understand both E2 and its receptor, the estrogen receptor (ER). The ER is a member of the nuclear receptor (NR) superfamily. The NR superfamily consists of a large group of proteins which regulate a large number of homeostatic proteins together with regulator proteins termed coregulators and corepressors. SRA (steroid receptor RNA activator) is the only known RNA coactivator and augments transactivation by NRs. SRA has been demonstrated to play an important role in mediating E2 action (Lanz et al., 1999; Lanz et al., 2003) and its expression is aberrant in many human breast tumors, suggesting a potential role in breast tumorigenesis (Murphy et al., 2000). Despite evidence that an alternative splice variant of SRA exists as a protein (Chooniedass-Kothari et al., 2004), it has been conclusively shown that SRA can function as an RNA transcript to coactivate NR transcription (Lanz et al., 1999; Lanz et al., 2002; Lanz et al., 2003). The precise mechanism by which SRA augments ER activity remains unknown. However, it is currently hypothesized that SRA acts as an RNA scaffold for other coregulators at the transcription initiation site. Several SRA stem loops have been identified as important for SRA function, including structure (STR) 1, 5 and 7 (Lanz et al., 2002; Zhao et al., 2007). Previously, I sought to identify SRA-binding proteins using a specific stem-loop structure of SRA (STR7) that was identified as both important for its coactivator function (Lanz et al., 2002) and also as a target for proteins from breast cancer cell extracts (Hatchell, 2002). From a yeast E. Hatchell Abstract iii III hybrid screen using STR7 as bait, I identified a novel protein which was named SLIRP (Patent Number: WO/2007/009194): SRA stem-Loop Interacting RNA-binding Protein (Hatchell, 2002; Hatchell et al., 2006). '...' This thesis demonstrates that SLIRP modulates NR transactivation, provides mechanistic insight into interactions between SRA, SRC-1, HSP-60 and NCoR and suggests that SLIRP may regulate mitochondrial function. These studies contribute significantly to the growing field of NR biology, and contribute more specifically to the elucidation of estrogen action in breast cancer. Furthermore, it lays a strong and exciting foundation for further studies to evaluate SLIRP as a biomarker and potential therapeutic target in hormone dependent cancers.

Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed Feb. 7, 2008). Includes bibliographical references.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2005. / Härtill 5 uppsatser.

Fabrication of effective algae cultivation systems adjacent to coal-fired power plants to fixate waste CO2 would represent a sizable step towards achieving a carbon neutral energy cycle. However, emission gas would elevate the algal cultivation system temperature and decreases its pH without expensive preprocessing. Increased temperature and acidity constitutes a profound stress on the algae. Although stressed algae produce heat shock proteins (HSPs) that promote protein folding and protect against stress, the ordinary biological response is insufficient to protect against coal flue gas. Experimental upregulation of HSPs could make algae respond to the stress caused by high temperatures and low pH at an elevated level. However, no work has been done to determine whether HSPs can be experimentally upregulated in algae. Here, the Chlamydomonas reinhardtii algal strain was selected because it has a sequenced genome and singular cell structure ideal for genetic modifications. Two genetic modification methods: transformation with plasmids pCB720/pCB740, and cloned pchlamiRNA3/pchlamiRNA3int with yeast HSP gene SSA1 were evaluated. pCB720/pCB740 up regulate algae production of native HSP, HSP70B. pCB720 transformation success was observed but statistically, data varied. pchlamiRNA3/pchlamiRNA3int were cloned with SSA1. Chlorophyll content measured growth indirectly. Quantitative HSP detection could be done using RT-PCR.

Submitted by LUÍS FELIPE DAIBES DE ANDRADE null (luipedaibes@hotmail.com) on 2017-12-09T19:13:02Z No. of bitstreams: 1 TeseVersaoFinal_LFD.pdf: 4542971 bytes, checksum: dc820c668f873e34980831a5f5f98979 (MD5) / Approved for entry into archive by Adriana Aparecida Puerta null (dripuerta@rc.unesp.br) on 2017-12-11T16:56:37Z (GMT) No. of bitstreams: 1 andrade_lfd_dr_rcla.pdf: 4542971 bytes, checksum: dc820c668f873e34980831a5f5f98979 (MD5) / Made available in DSpace on 2017-12-11T16:56:37Z (GMT). No. of bitstreams: 1 andrade_lfd_dr_rcla.pdf: 4542971 bytes, checksum: dc820c668f873e34980831a5f5f98979 (MD5) Previous issue date: 2017-12-01 / Outra / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O fogo é o principal distúrbio em diversas vegetações ao redor do mundo, denominadas ecossistemas inflamáveis. Neste contexto, muitas espécies possuem estratégias de regeneração e colonização do ambiente pós-fogo, o que tipicamente envolve a sobrevivência (tolerância) das sementes e/ou quebra da dormência. Em especial, muitas sementes de leguminosas possuem o tegumento impermeável (dormência física), que pode ser rompido por meio de choques térmicos relacionados ao fogo. Outro fator que pode auxiliar no processo de quebra da dormência física é a flutuação térmica no solo, cuja amplitude é aumentada nas clareiras formadas pela remoção da vegetação durante a queima. Ambos os fatores, fogo e flutuação térmica, são relativamente bem estudados nos ecossistemas inflamáveis da Austrália e em vegetações Mediterrâneas. Por outro lado, os padrões relacionados à quebra da dormência e germinação das sementes ainda são controversos e menos conhecidos nas savanas tropicais da África e da América do Sul. Assim, esta Tese de doutorado teve como objetivo avaliar o papel do fogo na germinação e sobrevivência das sementes de leguminosas do Cerrado. Para tanto, realizamos tratamentos em campo, incluindo queimas experimentais, e também aplicamos tratamentos no laboratório, simulando a flutuação térmica nas clareiras e a passagem do fogo (choques térmicos). Após os tratamentos, observamos a germinação em condições ótimas, fazendo contagens três vezes por semana, e realizamos testes de viabilidade ao final de 30 dias dos experimentos. As análises estatísticas consistiram basicamente em GLMMs com distribuição binomial, considerando as réplicas como efeitos aleatórios. Os resultados apontam que as sementes morrem quando diretamente expostas ao fogo na superfície do solo. Por outro lado, há uma maior probabilidade de sobrevivência quando as sementes estão localizadas em clareiras (gaps) da vegetação. Nos gaps, a maior porcentagem de solo nu proporciona temperaturas do fogo mais amenas, queimando por menos tempo. Quando enterradas 1-cm sob o solo, as sementes sempre sobrevivem e pode haver quebra de dormência. A flutuação térmica também pode quebrar uma proporção significativa da dormência em condições de campo, especialmente em Mimosa leiocephala. Em laboratório, ao contrário do esperado, não há quebra da dormência, indicando que a flutuação térmica em si não consiste um mecanismo para quebra da dormência física em espécies do Cerrado. Nos choques térmicos, poucas espécies (seis de 46) apresentaram quebra da dormência. Observamos também que a mortalidade das sementes está relacionada a um trade-off entre forma de crescimento, tamanho das sementes e presença de dormência. Isso se deve ao tamanho pequeno das sementes de arbustos, que a despeito de tipicamente ocorrerem em savana aberta, podem morrer sob condições severas dos choques térmicos, mais do que as espécies arbóreas. Dentre as espécies arbóreas, a filogenia influencia no tamanho e, consequentemente, na mortalidade das sementes frente ao fogo. Concluímos que a germinação das sementes não está diretamente ligada à presença do fogo no mosaico Cerrado-floresta, contrastando com os padrões reconhecidos para outros ecossistemas inflamáveis. / Fire is the principal disturbance in several vegetation types around the world, being called flammable ecosystems. In this context, many species have strategies to regenerate and colonize the post-fire environment, which typically involve seed survival (tolerance) and/or breaking of dormancy. In special, legume seeds usually have an impermeable seed coat (physical dormancy), which may be disrupted by fire-related heat shocks. Another factor that can help in the process of dormancy-breaking is the temperature fluctuation in the soil, which amplitude is increased in gaps formed by the removal of vegetation as a result of the passage of fire. Both factors, fire and temperature fluctuation, are relatively well studied in flammable ecosystems from Australia and Mediterranean vegetation. On the other hand, the patterns related to dormancy-breaking and seed germination are still controversial and less known in tropical savannas from Africa and South America. Therefore, this doctoral Thesis has aimed to evaluate the role of fire on germination and survival of legume seeds from Cerrado. Therefore, we conducted experiments in the field, including experimental burning, and also applied treatments in the laboratory, simulating temperature fluctuation in the gaps and fire passage (heat shocks). After the treatments, we observed germination under optimal conditions, making counting three times a week, and accomplishing viability tests by the end of 30 days of the experiment. The statistical analyses basically consisted on GLMMS with binomial distribution, considering replicates as random effects. Results pointed that seeds die when directly exposed to fire in the soil surface. On the other hand, there is a higher probability of survival when seeds are placed in vegetation gaps. In the gaps, the higher percentage of bare soil provides milder fire temperatures, lasting for less time. When buried at 1-cm belowground, seeds always survive and dormancy-breaking may occur. Temperature fluctuation can also break a significant proportion of dormancy under field conditions, especially for Mimosa leiocephala. In the laboratory, there is no dormancy-breaking, showing that temperature fluctuation itself might not be a mechanism for physical dormancy-breaking in Cerrado species. When heat shocks were applied, six out of 46 species showed dormancy-breaking. We also observed that seed mortality is related to a trade-off between growth-form, seed size and the presence of dormancy. This is due to the small size of seeds from shrubs, that although typically occurring in the open savannas, they might die under severe conditions of temperatures. Among the tree species, phylogeny influences on size and, consequently, mortality of seeds in face to fire. We concluded that seed germination is not directly linked to the presence of fire in the Cerrado-forest mosaic, contrasting to the patterns recognized for other flammable ecosystems. / Fundação Grupo Boticário 0153_2011_PR / FAPESP 2015/06743-0 / CAPES/PDSE 88881.131702/2016-01 / CNPq 455183/2014-7

Advances in next-generation sequencing platforms during the past decade have resulted in exponential increases in biological data generation. Besides applications in determining the sequences of genomes and other DNA elements, these platforms have allowed the characterization of cell-wide mRNA pools under different conditions and in different tissues. In 2009, Ingolia and colleagues developed an extension of high-throughput sequencing that provides a snapshot of all cellular mRNA fragments protected by translating ribosomes, dubbed ribosome profiling. This approach allows detection of differential translation activity, annotation of novel protein coding sequences and variants, identification of ribosome pause sites and estimates of de novo protein synthesis. As with other sequencing based methodologies, a major challenge of ribosome profiling has been sorting, filtering and interpreting the gigabytes of data produced during the course of a typical experiment. In this thesis, I developed and applied computational pipelines to interrogate ribosome profiling data in relation to gene expression in several viruses and eukaryotic species, as well as to identify sites of ribosomal pausing and sites of non-canonical translation activity. Specifically, I applied various control analyses for characterizing the quality of profiling data and developed scripts for visualizing genome-based (exon-by-exon) rather than transcript-based ribosome footprint alignments. I also examined the challenge of mapping footprints to repetitive sequences in the genome and propose ways to mitigate the associated problems. I performed differential expression analyses on data from coronavirus-infected murine cells, retrovirus-infected human cells and temperature-stressed Arabidopsis thaliana plants. Dissection of translational responses in Arabidopsis thaliana during heat shock or cold shock revealed several groups of genes that were highly upregulated within 10 minutes of temperature challenge. Analysis of the branches of the unfolded protein and integrated stress responses during coronavirus infection allowed for deconvolution of transcriptional and translational contributions. During the course of these analyses, I identified errors in a recently publicized algorithm for detection of differential translation, and wrote corrections that have now been pulled into the repository for this package. Comparison of the translational kinetics of the dengue virus infection in mosquito and human cell lines revealed host-specific sites of ribosome pausing and RNA accumulation. Analysis of HIV profiling data revealed footprint peaks which were in agreement with previously proposed models of peptide or RNA mediated ribosome stalling. I also developed a simulation to identify transcripts that are prone to generating RPFs with multiple alignments during the read mapping process. Together, the scripts and pipelines developed during the course of this work will serve to expedite future analyses of ribosome profiling data, and the results will inform future studies of several important pathogens and temperature stress in plants.

Submitted by Warne Pedro de Andrade (warne800@hotmail.com) on 2018-06-27T16:37:27Z No. of bitstreams: 1 Mestrado Warne.pdf: 2077163 bytes, checksum: 68e5205b57ffe2680fcdd40ebea34554 (MD5) / Approved for entry into archive by ROSANGELA APARECIDA LOBO null (rosangelalobo@btu.unesp.br) on 2018-06-28T16:07:28Z (GMT) No. of bitstreams: 1 andrade_wp_me_bot.pdf: 2077163 bytes, checksum: 68e5205b57ffe2680fcdd40ebea34554 (MD5) / Made available in DSpace on 2018-06-28T16:07:28Z (GMT). No. of bitstreams: 1 andrade_wp_me_bot.pdf: 2077163 bytes, checksum: 68e5205b57ffe2680fcdd40ebea34554 (MD5) Previous issue date: 2018-02-28 / Introdução: As proteínas de choque térmico (“Heat Shock Proteins”) são produzidas em resposta ao estresse patofisiológico nas células animais e não só fazem parte de várias etapas da carcinogênese, atuando principalmente como agentes antiapoptóticos, como também estão implicadas em mecanismos de resistência à quimioterapia em vários tipos de tumores. Objetivo: O presente estudo visa comparar a expressão dos genes TRAP1, HSPB1, HSPD1, HSPA1L e HSPA1A nas amostras de CEO (no tumor primário ou na metástase) com a expressão dos mesmos em amostras de tumores ovarianos benignos e tecido ovariano normal e correlacionar a expressão gênica com o prognóstico das pacientes e com a resistência ao tratamento com platina. Métodos: Foram avaliadas amostras de 51 pacientes operadas no Hospital Vera Cruz, entre os anos de 2008 a 2011, divididas em quatro grupos: CEO primário (n = 14), CEO metastático (n = 11), cistoadenoma seroso ovariano (n = 07) e ovário normal (n = 19). Utilizou-se a técnica de qRT-PCR para determinar o perfil de expressão dos genes. Resultados: As pacientes incluídas neste estudo apresentavam idade média de 56,75 anos. Não houve diferença significativa (valor-P> 0,050) na comparação entre a expressão dos genes e os grupos estudados. Os genes HSPA1A, HSPA1L e TRAP1 foram subexpressos e se diferiram significativamente dos genes em indivíduos com ovário normal. A expressão dos genes analisados não correlacionou se com as variáveis quantitativas, como idade, menarca, e tempo após menopausa bem como em relação as dosagens séricas do CA125, presença de ascite e citorredução. Em relação ao estadiamento, observou que mulheres com CEO no estadio I e II apresentaram a expressão do gene TRAP1 significativamente maior que as pacientes no estadio III e IV (p=0,040). Ao analisar a expressão dos genes em relação a sobrevida global observou se que houve influência significativa do nível de expressão dos gene HSPA1A sobre o risco de morte das mulheres afetadas por CEO. Conclusão: Apesar das limitações em relação ao tamanho da amostra, os resultados obtidos sinalizam para um melhor entendimento da relação entre a expressão destes genes e o câncer de ovário, evidenciando algumas implicações no prognóstico. / Heat Shock Proteins are produced in response to pathophysiological stress and take part in several stages of carcinogenesis, acting primarily as anti-apoptotic agents. They are also implicated in resistance to chemotherapy in several types of tumors. Herein we correlated the expression of genes encoding these proteins and the clinical and pathological aspects of patients with ovarian cancer (OC). METHODS: 51 patients included in the study were divided into four groups: those with primary EOC (n = 14), metastatic EOC (n = 11), ovarian serous cystadenoma (n = 7), with no evidence of ovarian malignancy or control (n = 11). The 57 tumor samples obtained were submitted to RNA extraction and reverse transcription. qRT-PCR was performed to compare the expression of TRAP1, HSPB1, HSPD1, HSPA1A and HSPA1L in primary and HSP60, HSP70, HSPA1L genes did not differ among the groups (p-value> 0.050) .HSPA1A, HSPA1L and TRAP1 we underexpressed in the primary and metastatic EOC groups with HSPA1L showing the lowest expression with compare with normal ovary tissue. TRAP1 expression was higher in tumors at stage I/II than at stages III/IV. Grade II subjects showed higher HSPB1 expression. There was no correlation between HSPs expression and age, menarche, parity, period after menopause initiation and CA-125. HSPA1A gene was negatively correlated with the risk of dying of OC. There was no differences between HSP expression gene evaluated and overall and disease-free survival. In conclusion, we suggest that downregulation of HSPA1A, HSPA1L e TRAP1 could help to clinical evaluated of prognostic in women with EOC.

The constitutively obsereved up-regulation of Hsp70 expression often led to premature conclusions about its critical role as a repair mechanism of cold injury that is, besides, expressed by protein misfolding/denaturation. In this study, we analyze the cold tolerance and the expression of 24 different mRNA transcripts of Hsp complex and other genes, that are associated with the repair of injury caused by cold. We use two strains of D. melanogaster: the wild type and the mutant type Hsp70- null, that lacks all 6 copies of the gene hsp70. We found out, that the larvae of two strains do not differ in their patterns of target genes expression during long term acclimation nor during recovery from chronic cold exposure and acute cold shock, therefore there is no transcriptional compensation of any other Hsp gene for the missing hsp70 in Hsp70- strain. The cold tolerance of Hsp70- strain larvae was impaired only, when they were exposed to strong acute cold shock by temperatures below -8°C. No difference in cold tolerance was observed, when the larvae were exposed to chronic cold exposure in 0°C or to mild acute cold shock by temperatures up to -4°C. Based on our results we assess, that the cold injury caused by strong acute cold schock is of another nature than caused by mild cold conditions and only in the first case Hsp70 expression is critical for the repair of cold injury in Drosophila melanogaster larvae.