Advances in Cell biology, vol. 32, 2005, issue 1

Anna GRĘDA, Danuta JANTAS

Abc of the ABC Transporters in Human Organism

Summary: Mitochondria play an essential role in the regulation of both cell survival and death. The irregularities of mitochondrial activity cause cells or all tissue dysfunction, can lead to insufficient energy supply. Consequently the mitochondrial perturbations can lead to cell death. In a broader spectrum of the mitochondrial dysfunction contribute to the pathogenesis of multiple neurodegenerative diseases including acute (e.g. ischemic stroke) and chronic disorders (Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, amyotrophic lateral sclerosis). Function of mitochondria can be in direct and/or indirect way interrupted by pathological factors, such as aggregates of misfolded mutant proteins (e.g. b-amyloid, tau protein, a-synuclein and mutant huntingtin), which affect cell functions and disrupt cell homeostasis. The main symptoms of mitochondrial dysfunction are: 1) insufficient energy supply and imbalance of ATP/ADP; 2) electron transport chain (ETC) activity reduction; 3) reactive oxygen species (ROS) overproduction; 4) Ca²⁺ overload; 5) release of pro-apoptic factors (e.g. cyt.c) and 6) deregulations in mitochondrial biogenesis process. Understanding the molecular mechanism of mitochondrial dysfunction in particular neurodegenerative diseases can lead to discovery of new therapeutic approaches (pharmacological and non-pharmacological) aimed to stop or delay neuronal degeneration.

Key words: mitochondrial apoptosis, neuroprotection, mitochondria permeability transition pore, caloric restriction, resveratrol, antioxidants

[Postępy Biologii Komórki 2012; 39: 321–344]

Anna KAZIENKO, Małgorzata PIASECKA, Anna RYMASZEWSKA, Dariusz GĄCZARZEWICZ, Rafał KURZAWA, Monika FRĄCZEK, Maciej KURPISZ, Maria LASZCZYŃSKA

Molecular Markers of Male Infertility: Nucleotide Polymorphisms of Sperm Chromatin Proteins – Part I

Summary: Testis-specific histones, protamines and transition proteins are involved in unique chromatin remodeling during differentiation of the spermatid. It is suggested that the protamines may have evolved from histone H1 ancestors. Probably, lysine-rich histones were conversed in to protamines – arginine and cysteine-rich proteins. Protamine 1 (P1) is present in all mammalian whereas protamine 2 (P2) only in some mammalian species. Genes for protamine (PRM1, PRM2) and transition protein 2 (TNP2) create common multigenic PRM1 → PRM2→TPN2 domain maps to chromosome 16 but gene locus for the transition protein 1 (TNP1) is located on chromosome 2. The occurrence of the genes in the common multigenic domain enables simultaneously gene expression. Moreover, the pseudogene (gene 4) called also protamine 3 is located in the domain between PRM2 and TNP2. In opposite to other genes of the domain product of gene 4 does not involve in sperm DNA compaction but probably plays role in sperm motility. TP1 protein is responsible for the removal of testis-specific histones from DNA, in turn TP2 takes part in the DNA packaging of differentiating spermatid. Nucleoproteine genes are composed of two exons and single intron. A single nucleotide polymorphism (SNP) are discovered in non-coding and coding regions in the nuclear protein genes. The synonymous and non synonymous substitutions are the effect of SNP in coding regions. Currently the studies indicate that polymorphism in the promoter region (c.-107G>C, c.-190C>A in PRM1) and 3’UTR (untranslated region, c.+62G>C in PRM2) of protamine genes and in the promoter region (c.-688A>T, deletion of 15 nucleotides -91 to -106 in TNP1) and intron (c.1030G>A in TNP2) of transition protein genes are the most important in non-coding regions. The substitution c.-107G>C in PRM1 was identified in oligozoospermic patients and in man with non-proven fertility, c.-190C>A in PRM1 was associated with reduced sperm morphology, c.+62G>C in PRM2 was found in subjects with low P1/P2 ratio. The polymorphism c.-688A>T and deletion of 15 nucleotides -91 to -106 in TNP1 and the substitution c.1030G>A in TNP2 were described in man with azoospermia. These polymorphic changes can result in the disturbances of the transcription and translation factor binding and finally can trigger protamine deficiency and lead to reduced male fertility. However the majority of identified substitutions do not play any role in the etiopathogenesis of male sterility. They constitute rare polimorphisms and they are discovered in fertile and infertile men with similar low frequency. The different comments concerning the significance of the SNPs can result from the population specific variants as well as from other unknown genetic changes associated with male infertility. Further research of new SNPs in nuclear protein genes that may serve as good molecular markers for genetic diagnosis of male infertility are suggested.

Key words: : spermatozoa, chromatin, DNA, protamines, SNP, polymorphism

[Postępy Biologii Komórki 2012; 39: 345–370]

Anna KAZIENKO, Małgorzata PIASECKA, Anna RYMASZEWSKA, Dariusz GĄCZARZEWICZ, Rafał KURZAWA, Monika FRĄCZEK, Maciej KURPISZ, Maria LASZCZYŃSKA

Molecular Markers of Male Infertility: Alternations of Sperm Chromatin Transcription And Translation – Part II

Summary: Male infertility can be associated with sperm chromatin abnormalities resulting from aberrant structure and amount of protamines. The changes concern not only abnormal nucleotide sequence in the protamine genes but also expression and translation of the proteins. Therefore the regulation of these processes is a target of many experimental studies which try to reveal molecular background of male sterility particularly that of idiopathic origin. The protamine genes appear to be transcribed in the meiotic spermatocytes as well as round spermatids and to be translated in elongating spermatids. Transcription is regulated via methylation of DNA and transcriptional factors that bind to the promoter region (TFIID, TFIIA) and to the polymerase II RNA (testis specific isoform ALF, TAFIIt). The amount and forms of transcriptional factors are different in germinal cells compare to somatic cells. Transcriptional factor – CREM is known to activate and suppress expression of major genes of spermatogenesis including genes for protamines. The factor is activated by cAMP or by ACT (LIM-domain family proteins, LIM-only protein) alternatively. Moreover nuclear matrix proteins play an essential role in the expression of described genes. The nuclear matrix bind to specific-haploid matrix attachment regions (MAR) flanking at both sides of multigenic PRM1→ PRM2→ TNP2 protamine domain that contain protamine 1, 2 and transition protein 2 loci, respectively. Nuclear adenylation of the protamine primary transcripts and regulatory proteins (PABP – Polyadenylate Binding Protein, family Y-box proteins) result in the stabilization of transcripts and inhibition of the translation. On the other hand, transcript deadenylation and removal of transcription/translation factors promote translation. Furthermore, the miRNAs binding to mRNA causes its cleavage or translation inhibition. Phosphorylated protamines are bound to specific lamine binding receptor (LBR) identified in nuclear fibrous lamina shortly after protamine synthesis and before binding to the DNA. In turn, the dephosphorylated protamines are liberated from LBR and progressively replace the transition proteins.

Key words: spermatozoa, chromatin, DNA, protamin, transcription, translation

[Postępy Biologii Komórki 2012; 39: 371–394]

Urszula KRASUSKA, Katarzyna BUDNICKA, Renata BOGATEK, Agnieszka GNIAZDOWSKA

Polyamines in Regulation of Seed Dormancy and Germination

Summary: Polyamines, mainly putrescine, spermine and spermidine play essential role in various physiological processes and in reaction to biotic and abiotic stresses, indicating their importance for plant survival. This review focus on polyamines action in seeds. In cooperation with classical phytohormones (abscisic acid, gibberellins, ethylene) and small signaling molecules, such as reactive oxygen species (ROS) or nitric oxide (NO), polyamines take part in regulation of embryogenesis, ageing, dormancy removal and germination of seeds. Polyamines are abundant in mature seeds, although the contribution of particular polyamines differ even among the same species. The effect of spermine in seed physiology, in general, contrast with other polyamines: putrescine and spermidine emphazing that individual polyamines have defined action in plants and that they differentially affect seed dormancy and germination. Manipulation of polyamine biosynthesis and/or catabolism leads to alteration in seedling development due to modification in seed embryogenesis resulting in seed abortion or restriction of germination.

Key words: dormancy, germination, polyamines, seeds, stratification

[Postępy Biologii Komórki 2012; 39: 395–414]

Iwona OSIŃSKA, Joanna DOMAGAŁA-KULAWIK

Procalcitonin in Acute Respiratory Infection

Summary: The procalcitonin (PCT) is a precursor peptide from the hormone calcitonin (CT). PCT is used in diagnosis and monitoring of bacterial infections. In severe infections, such as sepsis, the PCT may reach very high concentrations in human serum (even up to 1000 ng/ml). The normal range is below 0.1 ng/ml. Procalcitonin secretion occurs in the classical or alternative pathway and the alternative pathway serves an important role in bacterial infections. The study presents the available methods of determination procalcitonin, both quantitative tests and qualitative tests. Immunoluminometric assay (ILMA) is a reference test for all commercially available quantitative PCT test and the main stage of this test is the antigen-antibody reaction. The role of semi-qualitative test (POCT) was presented, which benefit is rapid diagnosis of infection and appropriate treatment. Very useful is also the fully automated test for the procalcitonin determination. The significance of procalcitonin in acute respiratory infection was discussed. The bacterial infection may lead to an exacerbation of chronic obstructive pulmonary disease and hospital-acquired pneumonia or community-acquired pneumonia with increasing concentrations of procalcitonin. The results of many studies show that PCT can be used as a prognostic marker in patients with ventilation associated pneumonia. The determination of PCT concentration seems to be important in patients with lung disease. It lets to exclude false-positive diagnosis of infection and serves as the criterion for the decision on the use antibiotic therapy.

Key words: procalcitonin, community-acquired pneumonia, bacterial infection, viral infection, COPD, ventilator-associated pneumonia, hospital-acquired pneumonia

[Postępy Biologii Komórki 2012; 39: 415–428]

Izabela TYLMAN, Stanisław KOWALCZYK

Receptors and Signaling Pathways Governing Root Nodule Symbiosis and Arbuscular Mycorrhiza

Summary: The symbiotic interactions between legumes and rhizobacteria are initiated by flavonoids exuded by the legume roots that in rhizobium species induce biosynthesis of lipo-chito-oligosaccharide signaling molecules, termed nodulation factors (Nod factors), which mediate the nodulation process in the receptive host. The Nod factor perception in legume roots causes curling of root hairs that entrap the rhizobacteria, leading to formation of infection thread and development of nitrogen-fixing root nodules. The Nod factor receptors, identified in four legume species (Lotus japonicus, Medicago truncatula, Glycine max, Pisum sativum), have a common structure composed of an extracellular lysine motif (LysM) anchored to a single-pass transmembrane domain and intracellular kinase domain, and are thus termed LysM receptor-like kinases (LysM-RLKs). The current model predicts two receptor-like kinases located on epidermal cells and infection thread that are involved in NF binding: in L. japonicus LjNFR1 and LjNFR5, in M. truncatula MtLYK3 and MtNFP, in G. max GmNFR1a/b and GmNFR5a/b, and in P. sativum PsSYM and PsSYM10. Another receptor-like kinase involved in symbiotic signaling has leucine rich repeat (LRR) and serine/threonine kinase domains and is encoded by LjSYMRK, MsNORK, MtDMI2, GmNORK and PsSYM19. Nod factor perception initiates a downstream signal transduction cascade. This involves three nucleoporins, encoded by LjNup133, LjNup85 and LjNENA, potassium ion-channel proteins localized in the nuclear membranes, encoded by LjCASTOR, LjPOLLUX and MtDMI1, as well as, a calcium and calmodulin-dependent protein kinase (CCaMK), encoded by MtDMI3/LjCCaMK/ PsSYM9. Following the perception of NF through LysM-RLK, biphasic Ca²⁺ signaling is induced in root hair cells, i.e. a rapid influx of Ca²⁺ into the hair cells cytosol and then periodical oscillation (Ca²⁺ spiking) of calcium concentrations within the nucleus and at perinuclear region. CCaMK, a putative decoder of Ca²⁺ signals, contains three functional domains, i.e. a serine/threonine-kinase domain, a calmodulin-binding (autoinhibitory) domain, and a domain of three EF-hands which interact with Ca²⁺ ions. Genetic and protein-protein interaction studies have identified protein components that interacts with CCaMK, and are required for NF signaling, namely Interacting Protein of DMI3 (MtIPD3), LjCYCLOPS and MtVapyrin. Moreover, several transcription factors activated downstream of CCaMK have been identified, including Nodulation Signaling Pathway1 and 2 (NSP1, NSP2), Ets2 Repressor Factor (ERF) required for nodulation (ERN1, ERN2 and ERN3) and Nodule Inception (NIN). Moreover, recent data strongly support a crucial role for cytokinins in promoting the activation and re-differentiation of cortical cells that initiate nodule development. Cytokinin receptor (LjLHK1/MtCRE1) and response regulators (RRs) are required for activation of the downstream transcription factors ERN1, NSP2 and NIN. Nodulation and the subsequent nitrogen fixation carry a metabolic cost to host plants, therefore to keep an appropriate nodule number, legumes possess regulatory system called “autoregulation of nodulation” (AON).
Over 80% of the angiosperm plants establish symbiotic relationships with mycorrhizal fungi that improve their ability to acquire nutrients from the soil. Evolutionary older arbuscular mycorrhiza (AM) is established between fungi belonging to the phylum Glomeromycota and the roots of most vascular plants. Despite the remarkable morphological differences between AM and root nodule symbioses, to date, at least nine genes have been identified in legume, the so-called common symbiosis (SYM) genes, that are required for the establishment of both fungal and bacterial symbioses.

Key words: Nod factor receptors, common symbiotic signaling pathway, nodule symbiosis, mycorrhizal symbiosis

[Postępy Biologii Komórki 2012; 39: 429–458]

Sylwia Olimpia RZOŃCA, Monika Ewa GOS

Role of FMRP Protein: Function and Role in Pathogenesis of Fragile X Syndrome

Summary: Fragile X syndrome (FXS) is the most common form of inherited X-linked intellectual disability and a leading known heritable cause of autism spectrum disorders. More than 99% of FXS cases are caused by CGG triplet expansion (>200 repeats) in 5’ untranslated region of the FMR1 gene that leads to low expression or absence of the FMRP protein. The FMRP is one of the key regulator protein involved in process of the translation in neural cells. It affects the peptide synthesis, the stabilization of the transcript factor and also is involved in the transport of mRNA to place of its target activity. The FMRP is particularly important for the process of synaptic plasticity (including long-term depression, LTD), and therefore for the mechanisms of learning and long term memory development. Studies in animal models of FXS, have given the start of the theory about the mGlu receptor dependent pathogenesis of fragile X syndrome. At present it is known that an abnormal synaptic plasticity is one of the fundamental causes of FRAX syndrome. The development of effective method of the treatment for FXS based on the mGluR5 receptor antagonists is a priority in the current research. Currently certain substances that inhibit mGLuR activity are testing in the clinical trials, such as the STX209, RO491753 or AFQ056. This work is a summary of the current state of knowledge concerning the FMRP protein structure, its activity and function as well as its role in the pathogenesis of fragile X syndrome and potential therapeutic strategies of this disease.

Keywords: intellectual disability, fragile X syndrome, FMRP protein, FMR1 gene

[Postępy Biologii Komórki 2012; 39: 459–476]

Ewelina STARZYŃSKA, Stanisław KOWALCZYK

Subcellular Relocation of PIN Proteins, and the Auxin Dependent Plant Growth and Development

Summary:Auxin movement between cells depends on AUX1/LAX, ABCB/PGP and PIN protein families that mediate auxin transport across the plasma membrane. The directionality of auxin flow within tissues is largely determined by polar, subcellular localization of PIN auxin efflux carriers that undergo rapid subcellular dynamics. At the cellular level, various internal and external signals are translated into specific changes in the polar, subcellular localization of PIN proteins thereby directing and redirecting the intercellular fluxes of auxin. These dynamic, rapid switches of PIN polarity are presumably enabled by permanently, repeating cycles of endocytosis and exocytosis of PIN proteins. The experimental data strongly suggest that phosphorylation and dephosphorylation of PIN transporters play an important role in determining the apical-basal polarity. The subcellular trafficking of PIN proteins appears to be regulated by ARF, RAB and ROP monomeric G proteins. Very recently, Auxin Binding Protein1 (ABP1) has been ascribed a role in regulation of endocytosis and ROP-mediated cytoskeletal rearrangements. Moreover, it has been revealed that the extracellular matrix is essential to maintain polar distribution of PIN proteins at the plasma membrane. Rapid PIN protein relocations have been observed during embryonic development. At least four PIN genes are dynamically expressed during embryogenesis. Immediately after the division of the zygote, PIN7 protein is localized to the apical side of the basal cell. At the 32-cell stage, PIN7 reverses to the basal membrane of the suspensor cells. PIN1 protein is expressed without any polarity in the embryo, however at the 32-cell stage, PIN1 transporter becomes polarly localized to the basal membranes in the provascular cells. The PIN4 protein is expressed in hyphophysis cell, and after division, in its uppermost daughter cell. The expression of PIN3 gene starts relatively late at the heart stage in the columella precursors. Shoot lateral organs (leaves and flowers) are generated from the apical meristem in a highly periodic phyllotactic pattern. In Arabidopsis thaliana, the 137º angle between developing primordial is marked by auxin maxima at the position of incipient primordial. Auxin arrives at the organ initiation sites through the epidermis layer and is canalized through the interior of developing primordial into the basipetal stream. PIN1 proteins polarity in the epidermis layer, as well as their complex rearrangements relative to auxin maxima appear to be responsible for generating the phyllotactic pattern of auxin distribution. A number of studies have shown that polar auxin transport is required for continuous vascular pattern formation and establishment of procambial strands. PIN1 transporter plays a major role in leaf vascular patterning because it is the only protein of the PIN family to be expressed in future vein cells. During the formation of vascular veins in leaves, PIN1 proteins direct auxin towards a convergence point in the leaf epidermis, from where veins are being initiated, and where PIN1 polar localization marks the position of all future veins. During postembryonic development, the activity of shoot and root apical meristems lead to the formation of shoot- and root-specific organs. In the axil of each leaf, one or more secondary axillary meristems are formed. The axillary meristem often initiates a few leaves before arresting their growth to form a dormant axillary bud. Polar auxin transport in stems is necessary for the control of bud outgrowth. Very recently, the experimental data strongly suggest that establishment of auxin export from the bud is crucial for the bud to be activated. This increased auxin export is accompanied by gradual PIN1 polarization in the activated buds. In addition, strigolactones reduce the accumulation of PIN1 proteins on the basal membrane of xylem parenchyma cells. Auxin transport is also crucial for lateral root initiation. In pericycle cells, auxin maxima specify the founder cells for lateral root initiation. During the initiation phase, PIN1 proteins are localized at the anticlinal membranes, and after the switch of the pericycle cell division plan, they redistributed to the outer lateral plasma membrane.

Key words: H Auxins, PIN proteins relocation, auxin and plant development

[Postępy Biologii Komórki 2012; 39: 477–502]

Natalia DERUS, Joanna FILIPOWSKA, Anna M. OSYCZKA

Multifunctional Germin Protein and Germin-Like Proteins in Plants

Summary: Mesenchymal Stem Cells (MSCs), especially those from bone marrow, are natural source of osteoblasts (bone forming cells). Therefore, many innovative experimental bone regeneration therapies are based on MSCs. Delivery of those cells into animal organisms often requires the use of appropriate growth surfaces or scaffolds that enable MSCs’ infiltration and support osteogenesis by those cells. This is possible thanks to surfaces and scaffolds enriched in silicon or silicon derivatives. The role of silica in osteogenesis is not fully understood yet. Silicon that is delivered to animal organisms mainly with food has a positive effect on the connective tissue, including e.g. the production of collagen in bone tissue and skin, and stimulation of cartilage extracellular matrix production. This review summarizes the latest reports regarding the application of silicon and its derivatives in surfaces and scaffolds, which are designated for osteogenic cells cultures, starting with bioactive ceramics and polymers and ending with metals and natural biomaterials. We present the most recent examples of silica-based materials influencing osteogenesis in vitro.

Key words: mesenchymal stem cells, osteoblasts, osteogenesis, silicon, surfaces and scaffolds for cell culture

[Postępy Biologii Komórki 20121; 39: 503–518]

Christopher KOBIERZYCKI, Agnieszka MALIŃSKA, Piotr DZIĘGIEL

Application of Tissue Microarray Technique (TMA) in Neoplastic Markers Studies on Breast Cancers

Summary:The tissue microarray technique (TMA) was presented for the first time over 15 years ago. Since then, its use grows steadily and preparations achieved with its help meet all standards required to conduct advanced research. In this article methods for creating classic TMA and the alternative solutions proposed by some authors will be shown. An overview of research conducted with the use of TMA will be presented, concerning studies for new markers of tumor cell proliferation in breast cancers. Moreover further possibilities and difficulties arising from the use of this modern method will be discussed. This article aims to draw attention to the possibility of more frequent use of tissue microarray technique for diagnostic and research conducted in Poland.

Key words: rtissue microarrays, TMA, proliferation markers, breast cancer

[Postępy Biologii Komórki 2012; 39: 519–530]

Małgorzata STATKIEWICZ, Maciej MAŁECKI

Role of Sonic Hedgehog Pathway in Carcinogenesis: Cancer Stem Cells, Multidrug Resistance, Angiogenesis

Summary: Sonic Hedgehog pathway plays an important role in the formation and development of many types of cancer. Its biological significance is not fully understood. Cancer stem cell theory explains how SHH may influence on metastasis and self-renewal of cancer cells. Furthermore, it was discovered a strong link between SHH and vascular endothelial growth factor VEGF, which is essential for vascularization, and thus in promoting tumor growth and metastasis. The constitutive activation of Sonic Hedgehog pathway also contributes to resistance of tumors to treatment with cytostatics. Preliminary studies indicate that the high activity of SHH pathway is correlated with an increased expression of ABC transporter membrane proteins and reduced sensitivity of tumor cells towards therapeutics.

Key words: Sonic hedgehog, cancer stem cells, multidrug resistance, angiogenesis

[Postępy Biologii Komórki 2012; 39: 531–553]