Molecular cloning and expression of two HSP70 genes in the prawn, Macrobrachium rosenbergii

Liu, Jun; Yang, Wei‐Jun; Zhu, Xiaojing; Karouna‐Renier, Natalie K.; Rao, Ranga

doi:10.1379/csc-40r.1

Cited by 77 publications

(44 citation statements)

References 36 publications

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“…In cells, the constitutive form Hsc70 remains unchanged (e.g., LeBlanc et al 2011) or is only slightly upregulated (up to 2-fold) in certain tissues (e.g., Liu et al 2004;Rendell et al 2006) whereas the inducible form Hsp70, is highly upregulated from low basal levels (Deane and Woo 2005). Thus, as both Hsp70 and Hsc70 confer thermal tolerance (e.g., Fangue et al 2006), accounting for both forms of Hsp70 is a better predictor of thermal tolerance (Sorte and Hofmann 2005).…”

Section: Hsp70 Extraction and Quantificationmentioning

confidence: 99%

Thermal tolerance of the crab Pachygrapsus marmoratus: intraspecific differences at a physiological (CTMax) and molecular level (Hsp70)

Madeira

Narciso

Cabral

et al. 2012

Cell Stress and Chaperones

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Temperature is one of the most important variables influencing organisms, especially in the intertidal zone. This work aimed to test physiological and molecular intraspecific differences in thermal tolerance of the crab Pachygrapsus marmoratus (Fabricius, 1787). The comparisons made focused on sex, size, and habitat (estuary and coast) differences. The physiological parameter was upper thermal limit, tested via the critical thermal maximum (CTMax) and the molecular parameter was total heat shock protein 70 (Hsp70 and Hsp70 plus Hsc70) production, quantified via an enzyme-linked imunosorbent assay. Results showed that CTMax values and Hsp70 production are higher in females probably due to different microhabitat use and potentially due to different hormonal regulation in males and females. Among females, non-reproducing ones showed a higher CTMax value, but no differences were found in Hsp70, even though reproducing females showed higher variability in Hsp70 amounts. As reproduction takes up a lot of energy, its allocation for other activities, including stress responses, is lower. Juveniles also showed higher CTMax and Hsp70 expression because they occur in greater shore heights and ageing leads to alterations in protein synthesis. Comparing estuarine and coastal crabs, no differences were found in CTMax but coastal crabs produce more Hsp70 than estuarine crabs because they occur in drier and hotter areas than estuarine ones, which occur in moister environments. This work shows the importance of addressing intraspecific differences in the stress response at different organizational levels. This study shows that these differences are key factors in stress research, climate research, and environmental monitoring.

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Section: Hsp70 Extraction and Quantificationmentioning

confidence: 99%

Thermal tolerance of the crab Pachygrapsus marmoratus: intraspecific differences at a physiological (CTMax) and molecular level (Hsp70)

Madeira

Narciso

Cabral

et al. 2012

Cell Stress and Chaperones

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“…When compared with homologous vent annelid sequences, the P. grasslei hsp70 form 1 is closely related to the Alvinella pompejana sequence, whereas the P. grasslei hsp70 form 2 is rather closer to the Hesiolyra bergi sequence. Several structural characteristics were proposed to differentiate hsc70 and hsp70, like the presence/absence of introns, respectively (Gunther and Walter, 1994;Boutet et al, 2003;Liu et al, 2004), or the occurrence of the motif GGMP in the C-terminal region for HSC70 (Prapapanich et al, 1996;Liu et al, 2004). However, according to Leignel and colleagues, since these criteria are specific for each organism, gene expression studies still seem to be the only way to distinguish hsp70 and hsc70 (Leignel et al, 2007).…”

Section: Relationship Amongst Annelid Hsp70 Proteinsmentioning

confidence: 99%

Thermal biology of the deep-sea vent annelidParalvinella grasslei:in vivostudies

Cottin

Ravaux

Léger

et al. 2008

Journal of Experimental Biology

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SUMMARYThe annelid Paralvinella grasslei is a deep-sea vent endemic species that colonizes the wall of active chimneys. We report here the first data on its thermal biology based on in vivo experiments in pressurized aquaria. Our results demonstrate that P. grasslei survives a 30 min exposure at 30°C, and suggest that the upper thermal limit of this species is slightly above this temperature. The first signs of stress were noticed at 30°C, such as a significant increase in the animal's activity and the expression of HSP70 stress proteins. A preliminary investigation of the kinetics of stress protein expression surprisingly showed high levels of HSP70 proteins as late as 3.5 h after the heat shock. Finally, we provide here the first sequences for vent annelid hsp70 (P. grasslei, Hesiolyra bergi and Alvinella pompejana). These constitute valuable tools for future studies on the thermal biology of these annelids.

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“…Ali et al 2003;Karouna-Renier et al 2003;Jiao et al 2004;Leignel et al 2007;Ming et al 2010). Also intronless hsc70 were reported (Liu et al 2004). Hence, the presence of two introns in the partial sequence belonging to the nucleotide-binding domain is conceivable, and more intervening sequences are likely to be present in the entire hsc70 gene.…”

Section: Resultsmentioning

confidence: 99%

An intron-containing, heat-inducible stress-70 gene in the millipede Tachypodoiulus niger (Julidae, Diplopoda)

Knigge

Bachmann

Köhler

2014

Cell Stress and Chaperones

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The highly conserved part of the nucleotidebinding domain of the hsp70 gene family was amplified from the soil diplopod Tachypodoiulus niger (Julidae, Diplopoda). Genomic DNA yielded 701, 549 and 540 bp sequences, whereas cDNA from heat shocked animals produced only one distinct fragment of 543 bp. The sequences could be classified as a 70 kDa heat shock protein (hsp70), the corresponding 70 kDa heat shock cognate (hsc70) and a glucoserelated hsp70 homologue (grp78). Comparisons of genomic and cDNA sequences of hsc70 identified two introns within the consensus sequence. Generally, stress-70 expression levels were low, which hampered successful RT-PCR and subsequent subcloning. Following experimental heat shock, however, the spliced hsc70 was amplified predominantly, instead of its inducible homologue hsp70. This finding suggests that microevolution in this soil-dwelling arthropod is directed towards low constitutive stress-70 levels and that the capacity for stress-70 induction presumably is limited. hsc70, albeit having introns, apparently is inducible and contributes to the stress-70 response.

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Molecular cloning and expression of two HSP70 genes in the prawn, Macrobrachium rosenbergii

Abstract: Two complementary deoxyribonucleic acid (cDNA) clones encoding 2 different 70-kDa heat shock proteins (HSPs) were isolated from the prawn Macrobrachium rosenbergii.

Cited by 77 publications

References 36 publications

Thermal tolerance of the crab Pachygrapsus marmoratus: intraspecific differences at a physiological (CTMax) and molecular level (Hsp70)

Thermal tolerance of the crab Pachygrapsus marmoratus: intraspecific differences at a physiological (CTMax) and molecular level (Hsp70)

Thermal biology of the deep-sea vent annelidParalvinella grasslei:in vivostudies

An intron-containing, heat-inducible stress-70 gene in the millipede Tachypodoiulus niger (Julidae, Diplopoda)

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