Ras association domain-containing protein 5 (RASSF5), one of the prospective biomarkers for tumors, generally plays a crucial role as a tumor suppressor. As deleterious effects can result from functional differences through SNPs, we sought to analyze the most deleterious SNPs of RASSF5 as well as predict the structural changes associated with the mutants that hamper the normal protein-protein interactions. We adopted both sequence and structure based approaches to analyze the SNPs of RASSF5 protein. We also analyzed the putative post translational modification sites as well as the altered protein-protein interactions that encompass various cascades of signals. Out of all the SNPs obtained from the NCBI database, only 25 were considered as highly deleterious by six in silico SNP prediction tools. Among them, upon analyzing the effect of these nsSNPs on the stability of the protein, we found 17 SNPs that decrease the stability. Significant deviation in the energy minimization score was observed in P350R, F321L, and R277W. Besides this, docking analysis confirmed that P350R, A319V, F321L, and R277W reduce the binding affinity of the protein with H-Ras, where P350R shows the most remarkable deviation. Protein-protein interaction analysis revealed that RASSF5 acts as a hub connecting two clusters consisting of 18 proteins and alteration in the RASSF5 may lead to disassociation of several signal cascades. Thus, based on these analyses, our study suggests that the reported functional SNPs may serve as potential targets for different proteomic studies, diagnosis and therapeutic interventions. Ras association domain-containing protein 5 (RASSF5) is the leading member of Ras effector super family protein that prevents tumor growth by facilitating G 1 /S arrest of cell cycle 1-6. RASSF5 is a proapoptotic component of Ras and induces p53 mediated apoptosis 4. RASSF5 is implicated in a range of cell reactions including apoptosis, senescence, cell cycle regulation, differentiation and cell proliferation 5. Inactivation of RASSF5 is found to be associated in the oncogenesis, proliferation and weak diagnosis of human cancers 5. In various tumor tissues, low RASSF5 expression has been discovered and can therefore be considered as a prospective biomarker for tumors. RASSF5, in a range of human cancers, is shown to play crucial functions and is identified as a putative tumor suppressor 7. The complete length of RASSF5 gene is 81 kb and it is located at the locus 1q32.1 that covers several isoforms resulting from alternative mRNA splicing and differential promoter usage, which may have different roles in oncogenesis 3. NORE1A, with a molecular weight of around 47 kDa, is a bigger isoform of the RASSF5A gene that encodes a protein of 418 amino acids 8. The RASSF5A protein structure includes an N-terminal proline-rich section containing prospective SH3-binding sites, and a nuclear localization signal, a cysteine-rich C1-type zinc finger (C1) domain, a Ras-association (RA) domain that interacts with GTP-bound H-Ras or several other Ras...

Bruguiera gymnorrhiza is an ecologically and somewhat economically important mangrove tree species. We isolated 10 polymorphic microsatellite loci from B. gymnorrhiza using a dual‐suppression polymerase chain reaction (PCR) technique. These loci provided microsatellite markers with polymorphism of two to five alleles per locus within 216 individuals from nine natural populations of B. gymnorrhiza on Iriomote Island, the Sakishima Islands, Japan. The expected and observed heterozygosities ranged from 0.220 to 0.720 and from 0.104 to 0.447, respectively.

Rhizophora stylosa is an ecologically important mangrove tree species that is found in tropical and subtropical regions. We isolated five polymorphic microsatellite loci from R. stylosa using a dual-suppression-polymerase chain reaction technique. These loci provided microsatellite markers with polymorphism of four alleles in each locus for overall population. The mean expected heterozygosities ranged from 0.113 to 0.473.

Chloroplast microsatellite (cpSSR) markers were developed for three ecologically and economically important tree species in the mangrove family, Rhizophoraceae: Bruguiera gymnorrhiza, Kandelia candel, and Rhizophora stylosa. Noncoding regions of chloroplast DNA (cpDNA) from each species were separately amplified using universal chloroplast primers. Six, two, and three polymorphic cpSSR loci in B. gymnorrhiza, K. candel, and R. stylosa, respectively, were developed from amplified noncoding cpDNA regions. Characterization of 216, 156, and 253 individuals of B. gymnorrhiza, K. candel, and R. stylosa, respectively, collected from different natural mangrove populations (B. gymnorrhiza, 9; K. candel, 7; R. stylosa, 9) on Iriomote Island in Japan showed that these loci provide cpSSR markers with polymorphisms ranging from two to four alleles per locus and gene diversity between 0.027 and 0.480. These cpSSR markers will be useful for analyzing the maternal lineage distributions and population genetic structures of the three species. Several of these markers may also be useful in similar studies of other mangrove species.Keywords cpSSR Á Mangrove Á Maternal lineage Á Rhizophoraceae Bruguiera gymnorrhiza, Kandelia candel, and Rhizophora stylosa are ecologically and economically important tree species of a widely distributed mangrove family, the Rhizophoraceae. Ecologically, these three species serve as important components of the coastal mangrove ecosystem, as they protect shorelines by controlling coastal erosion and reducing the damaging effects of tidal surges and cyclonic storms in the coastal areas, and maintain the food web of coastal marine ecosystems. These species are important economically to communities living adjacent to mangroves, because of their use as timber and fuelwood and as raw materials for cottage industries. Information on population genetics and mating systems can provide effective guidelines for ecological management and conservation of these species. In the present study, chloroplast microsatellite (cpSSR) markers were isolated from B. gymnorrhiza, K. candel, and R. stylosa in order to investigate their maternal lineages, population genetic structures, and mating systems.Genomic DNA was extracted from silica gel-dried leaves using a modified cetyltrimethyl ammonium bromide (CTAB) method . cpSSR regions were isolated from B. gymnorrhiza, K. candel, and R. stylosa by sequencing the noncoding regions of chloroplast DNA (cpDNA) following the method of Lian et al. (2003). Briefly, the cpDNA fragments of each species were amplified by PCR using 11 universal chloroplast primer pairs developed by Taberlet et al. (1991), Demesure et al.

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