New Insights into Protamine-Like Component Organization in<i>Mytilus galloprovincialis'</i>Sperm Chromatin

Vassalli, Quirino Attilio; Caccavale, Filomena; Avagnano, S.; Murolo, Alessandra; Guerriero, Giulia; Fucci, Laura; Ausió, Juan; Piscopo, Marina

doi:10.1089/dna.2014.2631

Cited by 31 publications

(37 citation statements)

References 34 publications

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“…Mussels are in fact an ideal sentinel species as they have an excellent bioaccumulative affinity, as widely documented, and are sensitive to low concentrations of pollutants, including metals (Piscopo et al, 2016). Furthermore, our previous studies highlight a new insight into Mytilus galloprovincialis sperm chromatin organization (Vassalli et al, 2015), whose protein component is mainly constituted by three Protamine-like proteins (PL-proteins): PL-II (20%), PL-III (50%) and PL-IV (6%), that coexist with about 20% of somatic histones. PL-proteins are extremely rich in lysine and arginine and belong to sperm nuclear basic proteins (SNBP) that partially or totally replace somatic histones during spermatogenesis.…”

Section: Introductionmentioning

confidence: 65%

Protamine-like proteins analyses as emerging biotechnique for cadmium impact assessment on male mollusk Mytilus galloprovincialis (Lamarck 1819)

et al. 2018

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Here we report the industrial pollution effects due to cadmium on the reproductive health of Mytilus galloprovincialis. Mussels were removed from the biofouling of a Conatex panel after one year exposition at a polluted site near a disposal metallurgical factory. A high cadmium bioaccumulation was observed in the testis of mussels housed at the polluted site, with respect to a control site, as determined by inductively coupled plasma-mass spectrometry, along with a 10 fold increase in metallothionein 20 kDa gene (mt20) expression levels determined by qPCR. Furthermore, mussels transferred into laboratory tanks from the reference site, and exposed to 1.5, 5 and 10 µM CdCl, revealed a 1.7, 3.2 and 4.5 fold expression increase in the testis mt20, respectively, and a positive correlation with cadmium bioaccumulation was found. To evaluate a potential detrimental risk of such alterations on spermatozoa, we carried out electrophoretic analyses on their protamine-like proteins. As determined by AU-PAGE, after 1.5 µM CdCl exposure, protamine-like proteins also display major alterations with respect to those obtained after 5 and 10 µM CdCl exposure. All protamine-like proteins isolated from the polluted biofouling were in an aggregated form and displayed the same reduced DNA binding affinity of the protamine-like proteins obtained after 1.5 µM CdCl as demonstrated EMSA with sperm genomic DNA. Our results contribute to the studies concerning cadmium induced testis alterations and highlight protamine-like proteins' analysis as an emerging biotechnique for cadmium impact assessment on Mytilus galloprovincialis, for the sensitivity of the in vivo and in vitro changes of protamine-like proteins' state and their DNA binding affinity.

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Section: Introductionmentioning

confidence: 65%

Protamine-like proteins analyses as emerging biotechnique for cadmium impact assessment on male mollusk Mytilus galloprovincialis (Lamarck 1819)

et al. 2018

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“…Since we found differences in the electrophoretic pattern of the PL proteins extracted from the mussels exposed to some hyposalinity conditions, we wanted to prove if there were differences in their DNA binding affinity with respect to those obtained from mussels exposed to 35.9 psu condition. For this aim we performed electrophoretic mobility shift assay (EMSA), using pGEM3 DNA plasmid as a probe, under the same conditions described in Vassalli et al (). We found that PL proteins extracted from mussels exposed to control (35.9 psu) and to 17.1 psu conditions had the same DNA binding affinity.…”

Section: Resultsmentioning

confidence: 99%

“…They represent the 76% of the overall nuclear protein in M . galloprovincialis with PL‐III (11.3 kDa) present in the highest amount (50%) and PL‐II (14 kDa), and PL‐IV (6.5 kDa), which represent the 20% and 6%, respectively (Vassalli et al, ). Further, we have recently proposed the PL proteins both as antibacterial agents (Notariale et al, ) and as an emerging biomarker for cadmium impact assessment on male M .…”

Section: Introductionmentioning

confidence: 99%

“…These proteins are related to histone H1 and are rich in arginine and lysine (30-50 mol%;Fioretti et al, 2012;Piscopo et al, 2010). They represent the 76% of the overall nuclear protein in M. galloprovincialis with PL-III (11.3 kDa) present in the highest amount (50%) and PL-II (14 kDa), and PL-IV (6.5 kDa), which represent the 20% and 6%, respectively (Vassalli et al, 2015). Further, we have recently proposed the PL proteins both as antibacterial agents and as an emerging biomarker for cadmium impact assessment on male M. galloprovincialis (De Guglielmo et al, 2019;Piscopo, Notariale, & Troisi, 2017;Piscopo, Trifuoggi, Notariale, et al, 2018).…”

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Molecular effects on spermatozoa of Mytilus galloprovincialisexposed to hyposaline conditions

Lettieri

Maione

Ranauda

et al. 2019

Molecular Reproduction Devel

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Salinity represents a critical environmental and an ecological factor in the reproduction of marine species. As global climate changes and anthropogenic factors affect salinity, in this study, we have analyzed the responses of Mytilus galloprovincialis spermatozoa to hyposaline stress. We exposed mussels, in laboratory tanks, for 24 hr at 18°C to control (35.9 psu) and three hyposaline (17.1, 22.6, and 26.2 psu) conditions, and evaluated the expression of sperm hsp70 and protamine‐like proteins genes. Further we analyzed the electrophoretic pattern, the DNA binding and the release from sperm nuclei of protamine‐like proteins. For all experimental approaches used, the results obtained at 17.1 psu condition were very similar to those obtained in the control condition, while alterations were always recorded at 22.6 and 26.2 psu conditions. Particularly, at 22.6 and 26.2 psu, was observed: 42.5‐ and 17.1‐fold increase in hsp70 expression, respectively, and hypoexpression of PL‐II/PLIV protamine‐like proteins genes. Further, electrophoretic mobility shift assays and salt‐induced release of nuclear proteins from sperm nuclei, revealed alterations in the PL proteins/DNA binding, in these two hyposaline conditions. The similarity between the results obtained in control and in the more severe hyposaline condition (17.1 psu) could indicate a phenomenon of fertility preservation strategy due to gamete plasticity.

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“…The crude extract containing the three PL-proteins from M. galloprovincialis spermatozoa was obtained by extraction with 5% PCA as previously described (Piscopo et al, 2018c), starting from a pool of spermatozoa collected from 20 Mytilus galloprovincialis specimens. M. galloprovincialis PL-II and PL-III were fractionated by ionic exchange resin cellulose Whatman CM52 as previously described (Vassalli et al, 2015). The lyophilized crude extracts containing the three PL-proteins and the single purified PL-proteins were stored at -80° C.…”

Section: Methodsmentioning

confidence: 99%

Protamine-like proteins have bactericidal activity. The first evidence in Mytilus galloprovincialis

et al. 2018

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The major acid-soluble protein components of the mussel Mytilus galloprovincialis sperm chromatin consist of the protamine-like proteins PL-II, PL-III and PL-IV, an intermediate group of sperm nuclear basic proteins between histones and protamines. The aim of this study was to investigate the bactericidal activity of these proteins since, to date, there are reports on bactericidal activity of protamines and histones, but not on protamine-like proteins. We tested the bactericidal activity of these proteins against Gram-positive bacteria: Enterococcus faecalis and two different strains of Staphylococcus aureus, as well as Gram-negative bacteria: Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella typhmurium, Enterobacter aerogenes, Enterobacter cloacae, and Escherichia coli. Clinical isolates of the same bacterial species were also used to compare their sensitivity to these proteins. The results show that Mytilus galloprovincialis protamine-like proteins exhibited bactericidal activity against all bacterial strains tested with different minimum bactericidal concentration values, ranging from 15.7 to 250 µg/mL. Furthermore, these proteins were active against some bacterial strains tested that are resistant to conventional antibiotics. These proteins showed very low toxicity as judged by red blood cell lysis and viability MTT assays and seem to act both at the membrane level and within the bacterial cell. We also tested the bactericidal activity of the product obtained from an in vitro model of gastrointestinal digestion of protamine-like proteins on a Gram-positive and a Gram-negative strain, and obtained the same results with respect to undigested protamine-like proteins on the Gram-positive bacterium. These results provide the first evidence of bactericidal activity of protamine-like-proteins.

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New Insights into Protamine-Like Component Organization inMytilus galloprovincialis'Sperm Chromatin

Cited by 31 publications

References 34 publications

Protamine-like proteins analyses as emerging biotechnique for cadmium impact assessment on male mollusk Mytilus galloprovincialis (Lamarck 1819)

Protamine-like proteins analyses as emerging biotechnique for cadmium impact assessment on male mollusk Mytilus galloprovincialis (Lamarck 1819)

Molecular effects on spermatozoa of Mytilus galloprovincialisexposed to hyposaline conditions

Protamine-like proteins have bactericidal activity. The first evidence in Mytilus galloprovincialis

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