Importin β-dependent nuclear import of TopBP1 in ATR–Chk1

checkpoint in Xenopus egg extracts and Evolution of a

cellular immune response in Drosophila:

a phenotypic and genomic comparative analysis.

Virginia Espinosa NavarroMedicine student

III SemesterUPB

Virginia Espinosa NavarroMedicine student

III SemesterUPB

BIBLIOGRAPHY

• Importin β-dependent nuclear import of TopBP1 in ATR–Chk1 checkpoint in Xenopus egg extracts. January 9, 2014. From: www.elsevier.com/locate/cellsig.

• Evolution of a cellular immune response in Drosophila: a phenotypic and genomic comparative analysis. Janyary 18, 2014. From: http://gbe.oxfordjournals.org/.

Importin β-dependent nuclear import of TopBP1 in

ATR–Chk1checkpoint in Xenopus egg extracts and Evolution of a

cellular immune response in Drosophila:

a phenotypic and genomic comparative analysis.

Teacher Lina Martínez

Virginia Espinosa Navarro

Medical studentIII semester

UPB

MEDICAL

UTILITY

This first study is of great clinical importance because, genetic and molecular biological studies offer us a path toward personalized treatment and prognostic factors, one of which includes the prevention of breast cancer, and it is known that family susceptibility to breast cancer comprises 25% of all breast tumors, so the ratio of the TopBP1 protein susceptibility to breast cancer is a step that leads us to find an appropriate and effective prevention.In the second study, the clinical importance of the phenotypic and genomic analysis comes from the possibility that the drug therapy can be safer and more effective when the genotype of the patient is known. Pharmacogenetic studies can be considered studies of interaction between genes and environment, considering pharmacological treatment as an environmental variable.

INTRODUCTIONImportin β-dependent

nuclear import of TopBP1 in ATR–Chk1

checkpoint in Xenopus egg extracts

Evolution of a cellular immune response in

Drosophila:a phenotypic and

genomic comparative analysis

In the first article, we seek to clarify one of the aspects that influence the susceptibility to breast cancer, and it’s importing the TopBP1 protein from the nucleus to the cytoplasm. Taking advantage of the cell-free Xenopus egg extract system, was investigated the roles of TopBP1 in DNA metabolism including DNA replication and DDR through a series of studies And all these data suggest that the Importin β-dependent shuttling of TopBP1 into nucleus plays an important role in the ATR–Chk1 checkpoint signaling in Xenopus.

At the second one, the propose that they had with the article was investigate the large interspecific differences in the ability to survive infection by parasitoids, across 11 Drosophila species and identify genomic changes associated with the evolution of cellular immunity in the Drosophila genu and the gains and losses of parasitoid resistance.

By identifying the importance of having the TopBP1 protein in DNA metabolism, replication, damage response and transcriptional regulation, and besides that, it has been identified that its location is associated with susceptibility to breast cancer , they wanted to do this study to investigate the way this core protein is transported to the cytoplasm, and a protein that interacted with TopBP1 C-terminal, known as B importin was found. After a series of procedures, managed to conclude important facts such as: Importin β has been identified as a new interacting protein of TopBP1 C-terminus.•A C-terminal motif in TopBP1 is important for its nuclear import.•Importazole inhibits TopBP1's nuclear import and ATR–Chk1 checkpoint signaling.•With ongoing DNA replication, nuclear import of TopBP1 is required for checkpoint.•Checkpoint activation requires more TopBP1 than DNA replication does.

As well the name says , this article seeks to evaluate the evolution of the immune response by phenotypic and genotypic comparison, for that, was investigated the large interspecific differences in the ability to survive infection by parasitoids across 11 Drosophila species and identify genomic changes associated with gains and losses of parasitoid resistance. They provide an important step towards understanding the evolution of the cellular resistance against parasitoids in Drosophila species and they showed that a combination of cooption and neofunctionalization is likely to have contributed to the acquiring of the new immunity component in the cellular immune response, and that particular gene families (serine-type proteases, Teps and lectins ) could be of special interest for the processes of hemocyte differentiation, proliferation and activation.

The susceptibility to breast cancer comprises 25% of all breast tumors, and the importing the TopBP1 protein from the nucleus to the cytoplasm plays an important role in this. Taking advantage of the cell-free Xenopus egg extract system, was investigated the roles of TopBP1 in DNA metabolism including DNA replication and DDR through a series of studies And all these data suggest that the Importin β-dependent shuttling of TopBP1 into nucleus plays an important role in the ATR–Chk1 checkpoint signaling in Xenopus.

The study about the evolution of a cellular immune response in Drosophila, show us that through a combination of phenotypic and genomic characterizations , they provide an important step towards understanding the evolution of the cellular resistance against parasitoids in Drosophila species. .

INT

RO

DU

CT

ION

Importin â-dependent nuclear import of TopBP1 in ATR–Chk1

checkpoint in Xenopus egg extracts

Xenopus egg extract preparation and related procedures

Recombinant proteins and antibodies

GST pulldown assays

Coimmunoprecipitation (Co-IP) assays

Immunofluorescence microscopy

MATERIALS AND METHODS

Importin β interacts with TopBP1

The CTM motif of TopBP1 mediates its interaction with Importin β

The CT100 fragment of Importin β is required for TopBP1 association

The CTM motif of TopBP1 is required for nuclear import from outside of assembled nuclei.

RESULTS

Importin β-specific inhibitor importazole compromised the nuclear import of TopBP1

The CTM-dependent nuclear import of TopBP1 is required for ATR–Chk1 checkpoint signaling, not due to its requirement for DNA replication

Nuclear import of TopBP1 plays a distinct regulatory role in the DNA damage response

RESULTS

OBSERVATION

Breast cancer is a reality that affects all women, and all of which are likely to have, it is vital to attend these studies, where molecular biology gives us significant events that influence the disease. In my opinion it is an article of great interest and it brings a lot to the medical science, indicating the influence of the TopBP1 protein in breast cancer susceptibility.

Evolution of a cellular immune response in

Drosophila:a phenotypic and genomic

comparative analysis

Species strains

Encapsulation assay

Lamellocyte identification

Melanization

Candidate genes

MATERIALS AND METHODS

Phylogenetic analysis

Immune classification

Positive selection

RT-qPCR

Orthologous groups and homology categories

MATERIALS AND METHODS

Phenotypic characterization: Only species of the melanogaster subgroup showencapsulation ability and produce lamellocytes

Comparative Genomics

Orthologs

Lineage specic gains and losses

Sequence divergence

Comparative expression of TepI, PPO3 and IM1

RESULTS

OBSERVATION

In this studio, they provide an important step towards understanding the evolution of the cellular resistance against parasitoids in Drosophila species, that’s important because, It has been argued that it may be beneficial to the public health use genetic information to stratify the allocation of environmental settings that prevent disease, although this view is not universally accepted.

MEDICAL

UTILITY

This first study is of great clinical importance because, genetic and molecular biological studies offer us a path toward personalized treatment and prognostic factors, one of which includes the prevention of breast cancer,.

MEDICAL

UTILITY

it is known that family susceptibility to breast cancer comprises 25% of all breast tumors, so the ratio of the TopBP1 protein susceptibility to breast cancer is a step that leads us to find an appropriate and effective prevention.

MEDICAL

UTILITY

In the second study, the clinical importance of the phenotypic and genomic analysis comes from the possibility that the drug therapy can be safer and more effective when the genotype of the patient is known. Pharmacogenetic studies can be considered studies of interaction between genes and environment, considering pharmacological treatment as an environmental variable

BIBLIOGRAPHY

• Importin β-dependent nuclear import of TopBP1 in ATR–Chk1 checkpoint in Xenopus egg extracts. January 9, 2014. From: www.elsevier.com/locate/cellsig.

• Evolution of a cellular immune response in Drosophila: a phenotypic and genomic comparative analysis. Janyary 18, 2014. From: http://gbe.oxfordjournals.org/.