Characterization of antifreeze protein gene expression in summer spruce budworm larvae

Khatiwada, Janak Raj; Tyshenko, Michael G.; Doucet, Daniel; Walker, Virginia K.

doi:10.1016/j.ibmb.2006.01.017

Cited by 12 publications

(5 citation statements)

References 29 publications

(42 reference statements)

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“…Even more striking, some transcripts appeared to peak in abundance in first instars and mRNA levels either did not change (CfAFP337) or decreased (CfAFP2.7a) during the overwintering stage when temperatures can drop as low as –30 °C. An intermediate 3′UTR transcript (CfAFP104) persisted to the third instar, a spring stage (Figs 1 and 4), and low levels of AFP mRNA were also detected in the summer, fifth instar stage (Qin et al ., 2006). In addition to this seemingly complex regulation, more than one transcript was often detected with the different probes, suggesting that different polyadenylation sites seen in the genomic clones are used in many of the different transcripts.…”

Section: Discussionmentioning

confidence: 99%

Transcription of antifreeze protein genes in Choristoneura fumiferana

Khatiwada

Doucet

Tyshenko

et al. 2007

Insect Molecular Biology

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Antifreeze proteins (AFPs) are encoded by approximately 17 genes in the spruce budworm, Choristoneura fumiferana. Northern analysis using 6 different cDNA probes showed isoform-specific patterns that varied during development. Transcripts for the majority of isoforms were most abundant in the second instar overwintering stage, but some were also detected in first instar and even in egg stages. In situ hybridization using riboprobes corresponding to two 9 kDa protein isoforms showed differential AFP expression even in second instars; CfAFP10 RNA was detected in all tissues, but CfAFP337 RNA distribution was more limited. Two genomic regions encoding three AFP genes have been isolated. Presumptive regulatory regions conferred transcriptional activity when placed upstream of a luciferase reporter sequence and transfected into a C. fumiferana cell line. The CfAFP2.26 core promoter is an 87 bp sequence containing a TATA box, whereas the CfAFP2.7 core promoter is a 76 bp sequence with both a TATA box and CAAT box, which directed higher reporter activities when tested in vitro. Reporter activity was not enhanced with five different hormones, although lower activities were observed with all intron-containing constructs. AFP message half-life, as assessed using reporter assays, was not appreciably influenced by isoform-specific-3'UTRs. These studies successfully demonstrate the temporal and spatial diversity of AFP expression encoded by this small gene family, and underscore the complexity of their regulation.

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Section: Discussionmentioning

confidence: 99%

Transcription of antifreeze protein genes in Choristoneura fumiferana

Khatiwada

Doucet

Tyshenko

et al. 2007

Insect Molecular Biology

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“…Other than these external environmental factors, many autoregulatory factors are also crucial in affecting the presence of AFP transcripts ( Qin and Walker 2006 ). For example, in C. fumiferana , AFP transcripts are most abundant in the second instar of the overwintering larvae, and are localized mainly in the fore and midguts of the larval body ( Qin et al 2006 ). In T. molitor , the presence of AFPs has been correlated with the stages of development ( Graham et al 2000 ).…”

Section: Discussionmentioning

confidence: 99%

“…Owing to the distinctive overwintering methods of different insects, the relationships between the presence and expression levels of AFPs and photoperiods, environmental temperatures, development stages, and the sexes of insects have been investigated ( Horwath et al 1996 , Graham et al 2000 ). The results of such multifaceted analyses indicate that insect AFPs contribute significantly to protecting insects against low temperatures in a variety of ways ( Qin et al 2006 ).…”

Section: Introductionmentioning

confidence: 99%

Sequencing and Expression Characterization of Antifreeze Protein Maxi-Like in Apis cerana cerana

Niu

Zhao

et al. 2018

Journal of Insect Science

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Antifreeze proteins (AFPs) are biological cryoprotectants with unique properties that play a crucial role in regulating the molecular mechanisms governing cold resistance in insects. To identify and characterize Apis cerana cerana AFP (AcerAFP), we cloned the full-length cDNA of AcerAFP and examined its expression patterns in A. cerana cerana. A nucleotide alignment analysis showed that the entire coding region of AcerAFP is 1095 bp and encodes a polypeptide of 365 amino acids. The amino acid sequence of this protein exhibits 63–96% homology with AFP homologs from other hymenopterans. α-helices form the main secondary and tertiary structures of AcerAFP, which is similar to the molecular structure of fish AFP type-I. The expression profiles of AcerAFP revealed that expression was tissue, sex, and developmentally specific. In response to cold stress, the mRNA and protein expression of AcerAFP were both induced by low temperatures, and were also related to the concentrations of several cryoprotectants, including glucose, glycerin, glutamic acid, cysteine, histidine, alanine, and methionine. In addition, we found that the knockdown of AcerAFP by RNA interference remarkably increased the total freezing temperature of hemolymph in A. cerana cerana, where levels of AcerAFP mRNA were correlated with the expression of most antifreeze-related proteins. Taken together, these results suggest that AcerAFP plays an essential role as a biological cryoprotectant in honeybees, and is in turn regulated by small cryoprotectants and antifreeze-related proteins.

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“…The protein structures of AFPs have been actively investigated to identify ice-binding motifs and determine ice controlling mechanisms [ 6 , 10 , 11 , 49 , 50 ]. However, little is known about the structure of AFPs from diatoms.…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of an Isoform Antifreeze Protein from the Antarctic Marine Diatom, Chaetoceros neogracile and Suggestion of the Core Region

et al. 2017

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Antifreeze proteins (AFPs) protecting the cells against freezing are produced in response to extremely low temperatures in diverse psychrophilic organisms, and they are encoded by multiple gene families. The AFP of Antarctic marine diatom Chaetoceros neogracile is reported in our previous research, but like other microalgae, was considered to probably have additional genes coding AFPs. In this paper, we reported the cloning and characterization of additional AFP gene from C. neogracile (Cn-isoAFP). Cn-isoAFP protein is 74.6% identical to the previously reported Cn-AFP. The promoter sequence of Cn-isoAFP contains environmental stress responsive elements for cold, thermal, and high light conditions. Cn-isoAFP transcription levels increased dramatically when cells were exposed to freezing (−20 °C), thermal (10 °C), or high light (600 μmol photon m−2 s−1) stresses. The thermal hysteresis (TH) activity of recombinant Cn-isoAFP was 0.8 °C at a protein concentration of 5 mg/mL. Results from homology modeling and TH activity analysis of site-directed mutant proteins elucidated AFP mechanism to be a result of flatness of B-face maintained via hydrophobic interactions.

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Characterization of antifreeze protein gene expression in summer spruce budworm larvae

Cited by 12 publications

References 29 publications

Transcription of antifreeze protein genes in Choristoneura fumiferana

Transcription of antifreeze protein genes in Choristoneura fumiferana

Sequencing and Expression Characterization of Antifreeze Protein Maxi-Like in Apis cerana cerana

Identification and Characterization of an Isoform Antifreeze Protein from the Antarctic Marine Diatom, Chaetoceros neogracile and Suggestion of the Core Region

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