Hormonal regulation of actin and tubulin in an epithelial cell line from Chironomus tentans

Fretz, A.; Spindler, K.-D.

doi:10.1002/(sici)1520-6327(1999)41:2<71::aid-arch3>3.3.co;2-9

Cited by 3 publications

(3 citation statements)

References 25 publications

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“…Actin fragments were amplified by using the primers Act U235 (AAYTGGGAYGAYATGGARAA) and Act L667 (GCCATYTCYTGYTCRAARTC) (Fretz & Spindler 1999) and the general actin primers Actup (ATG GTN GGN ATG GGN CAR AAR) and Actdown (ATG GTN GGN ATG GGN CAR AAR) (Viray Master, Chicago, 1993). PCR was run with 15 min initial denaturation at 94 °C followed by 35 cycles of 30 sec, denaturation at 94 °C, 1 min annealing at 50 °C, and 3 min elongation at 72 °C.…”

Section: Methodsmentioning

confidence: 99%

A cDNA library of the eutardigrade Hypsibius klebelsbergi Mihelčič, 1959 and analysis of the actin gene

Kiehl¹,

Dastych²,

D’Haese³

et al. 2007

J Limnol

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A cDNA library was constructed from the glacier-dwelling eutardigrade Hypsibius klebelsbergi from more than 2000 individuals collected in the Austrian Central Alps. RNA, DNA and proteins were successively isolated by the Trizol®-method. From the RNA preparation a cDNA library was constructed with the cDNA inserted unidirectionally in the phagemid expression vector TriplEx2. The primary gene library had a titre of 107 pfu ml-1 and the final amplified gene library a titre of 6×109 pfu ml-1. The average insert length was about 1.6 kb. The partial sequence of H. klebelsbergi actin (746 bp) showed highest similarity to GenBank data of Drosophila melanogaster actin at the nucleic acid level (84.9%) and at the amino acid level (98%). Compared with actin fragments of the eutardigrades Ramazzottius oberhaeuseri (450 bp) and Macrobiotus sp. (453 bp) the identities were 85% - 81% and 100% - 98% with respect to the nucleic/amino acids. Identity with actin fragments (359 bp) of Hypsibius dujardini from GenBank was 96% - 100%

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Section: Methodsmentioning

confidence: 99%

A cDNA library of the eutardigrade Hypsibius klebelsbergi Mihelčič, 1959 and analysis of the actin gene

Kiehl¹,

Dastych²,

D’Haese³

et al. 2007

J Limnol

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“…These results were in agreement with those obtained on mammalian and other insect cell lines and confirmed that the tubulin cytoskeleton was a target for steroid hormone-triggered changes in cell shape [26,31,32,44]. Other in vitro experiments performed on Drosophila Kc cells and mouse osteoblastic cells have revealed that steroid-induced morphological modifications were also accompanied by changes in the distribution and expression level of cytoskeleton actin [24,26,[45][46][47]. It would be interesting to determine whether steroid hormones act on these two major proteins of the cytoskeleton through a common signaling pathway.…”

Section: Discussionmentioning

confidence: 53%

Identification of steroid hormone signaling pathway in insect cell differentiation

Siaussat

Bozzolan

Porcheron

et al. 2007

Cell. Mol. Life Sci.

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To dissect the steroid hormone signaling pathway involved in insect cell morphological differentiation, we extended the application of the double-stranded RNA-mediated interference (dsRNAi) method to the epidermal IAL-PID2 cell line from Plodia interpunctella Lepidoptera. We first demonstrated that dsRNA was capable of efficiently blocking the steroid hormone 20-hydroxyecdysone (20E) inducibility of proteins that belong to the nuclear receptor superfamily, including the ecdysone receptor (EcR), its partner Ultraspiracle (USP), the insect homolog of the vertebrate retinoid X receptor and the HR3 transcription factor. We then showed that inhibiting the 20E induction of EcR, USP or HR3 proteins prevented the increased synthesis of beta tubulin and consequently the morphological transformation of cells. Thanks to this functional approach, we have shown, for the first time, the participation of EcR, USP and HR3 in a 20E signaling pathway that directs morphological differentiation in insect cells by regulating beta tubulin expression.

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“…2007) together with forward and reverse primers of conserved sequence regions of actin and specific primers of the expression vector λTriplEx2 (T 7, 5TripEx, 5Triplepostslip1). Actin fragments were amplified by using the primers Act U235 and Act L667 (Fretz and Spindler 1999). and the general actin primers Actup and Actdown (Viray master, Chicago 1993).…”

Section: Methodsmentioning

confidence: 99%

The actin gene of Hypsibius klebelsbergi (Eutardigrada) – complete sequence and comparison with actin from related and non‐related taxa

D’Haese

Traore‐Freitag

Kiehl

et al. 2011

Journal of Zoological Systematics and Evolutionary Research

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The actin gene of tardigrades was sequenced and analysed using a k ZAP Express cDNA library from the eutardigrade Hypsibius klebelsbergi previously constructed by us. We obtained the complete actin coding sequence of one isoform (1128 bp; 375 amino acids; MW 41 674 Da) together with parts of the 3¢-and 5¢-UTR region. Comparison of the 12 incomplete actin sequences of Hypsibius dujardini incorporated in GenBank indicates that this H. klebelsbergi actin sequence probably represents the most abundant muscle isoform. Ten of the H. dujardini clones show minor differences in codon usage and identical amino acid compositions to the H. klebelsbergi actin. Only two clones show amino acid variations in one and five positions, respectively, but show identical amino acids at their N-terminus. A considerable similarity between the 5¢-and 3¢-UTR regions of both tardigrade species was recognized. The H. klebelsbergi actin exhibits an overall high sequence similarity to the vertebrate b-actin. A comparison of muscle actins from various vertebrates as well as Ecdysozoa and non-Ecdysozoa revealed a more pronounced similarity of the tardigrade actin to arthropods and annelids and not to nematodes.

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Hormonal regulation of actin and tubulin in an epithelial cell line from Chironomus tentans

Cited by 3 publications

References 25 publications

A cDNA library of the eutardigrade Hypsibius klebelsbergi Mihelčič, 1959 and analysis of the actin gene

A cDNA library of the eutardigrade Hypsibius klebelsbergi Mihelčič, 1959 and analysis of the actin gene

Identification of steroid hormone signaling pathway in insect cell differentiation

The actin gene of Hypsibius klebelsbergi (Eutardigrada) – complete sequence and comparison with actin from related and non‐related taxa

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