TY  - JOUR
AB  - Mesenchymal stem cells (MSCs) are multilineage adult stem cells with considerable potential for cell-based regenerative therapies. In vitro expansion changes their epigenetic and cellular properties, with a poorly understood impact on DNA damage response (DDR) and genome stability. We report here results of a transcriptome-based pathway analysis of in vitro-expanded human bone marrow-derived mesenchymal stem cell (hBM-MSCs), supplemented with cellular assays focusing on DNA double-strand break (DSB) repair. Gene pathways affected by in vitro aging were mapped using gene ontology, KEGG, and GSEA, and were found to involve DNA repair, homologous recombination (HR), cell cycle control, and chromosomal replication. Assays for the recognition (gamma-H2AX + 53BP1 foci) and repair (pBRCA1 + gamma-H2AX foci) of X-ray-induced DNA DSBs in hBM-MSCs show that over a period of 8 weeks of in vitro aging (i.e., about 10 doubling times), cells exhibit a reduced DDR and a higher fraction of residual DNA damage. Furthermore, a distinct subpopulation of cells with impaired DNA DSB recognition was observed. Several genes that participate in DNA repair by HR (e.g., Rad51, Rad54, BRCA1) show a 2.3- to fourfold reduction of their mRNA expression by qRT-PCR. We conclude that the in vitro expansion of hMSCs can lead to aging-related impairment of the recognition and repair of DNA breaks.
AU  - Bao, X.
AU  - Wang, J.
AU  - Zhou, G.*
AU  - Aszodi, A.*
AU  - Schönitzer, V.*
AU  - Scherthan, H.*
AU  - Atkinson, M.J.
AU  - Rosemann, M.
C1  - 58945
C2  - 48491
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
SP  - 1238-1250
TI  - Extended in vitro culture of primary human mesenchymal stem cells downregulates Brca1-related genes and impairs DNA double-strand break recognition.
JO  - FEBS Open Bio
VL  - 10
IS  - 7
PB  - Wiley
PY  - 2020
SN  - 2211-5463
ER  - 

TY  - JOUR
AU  - Napolitano, V.*
AU  - Fino, R.
AU  - Softley, C.
AU  - Marciniak, M.*
AU  - Kalel, V.C.*
AU  - Mroz, P.*
AU  - Olmos, J.J.*
AU  - Blat, A.*
AU  - Davidowski, M.*
AU  - Erdmann, R.*
AU  - Popowicz, G.M.
AU  - Sattler, M.
AU  - Dubin, G.*
C1  - 57140
C2  - 47557
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
SP  - 426-427
TI  - Glycosomal protein import: A new target against trypanosomiasis.
JO  - FEBS Open Bio
VL  - 9
PB  - Wiley
PY  - 2019
SN  - 2211-5463
ER  - 

TY  - JOUR
AU  - Wątor, E.*
AU  - Zak, K.M.
AU  - Kalińska, M.*
AU  - Kuśka, K.*
AU  - Dubin, G.*
AU  - Popowicz, G.M.
AU  - Grudnik, P.*
C1  - 57141
C2  - 47556
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
SP  - 276-276
TI  - Crystal structure of a unique glucokinase from Kluyveromyces lactis.
JO  - FEBS Open Bio
VL  - 9
PB  - Wiley
PY  - 2019
SN  - 2211-5463
ER  - 

TY  - JOUR
AB  - Most lipases possess a lid domain above the catalytic site that is responsible for their activation. Lipase SMG1 from Malassezia globose CBS 7966 (Malassezia globosa LIP1), is a mono- and diacylglycerol lipase with an atypical loop-like lid domain. Activation of SMG1 was proposed to be solely through a gating mechanism involving two residues (F278 and N102). However, through disulfide bond cross-linking of the lid, this study shows that full activation also requires mobility of the lid domain, contrary to a previous proposal. The newly introduced disulfide bond makes lipase SMG1 eligible as a ratiometric thiol/disulfide redox potential probe, when it is coupled with chromogenic substrates. This redox-switch lipase could also be of potential use in cascade biocatalysis.
AU  - Guo, S.*
AU  - Popowicz, G.M.
AU  - Li, D.*
AU  - Yuan, D.*
AU  - Wang, Y.*
C1  - 48500
C2  - 41150
CY  - Hoboken
SP  - 477-483
TI  - Lid mobility in lipase SMG1 validated using a thiol/disulfide redox potential probe.
JO  - FEBS Open Bio
VL  - 6
IS  - 5
PB  - Wiley-blackwell
PY  - 2016
SN  - 2211-5463
ER  - 

TY  - JOUR
AB  - The use of TALEN and CRISPR/CAS nucleases is becoming increasingly popular as a means to edit single target sites in one-cell mouse embryos. Nevertheless, an area that has received less attention concerns the engineering of structural genome variants and the necessary religation of two distant double-strand breaks. Herein, we applied pairs of TALEN or sgRNAs and Cas9 to create deletions in the Rab38 gene. We found that the deletion of 3.2 or 9.3. kb, but not of 30. kb, occurs at a frequency of 6-37%. This is sufficient for the direct production of mutants by embryo microinjection. Therefore, deletions up to ~10. kb can be readily achieved for modeling human disease alleles. This work represents an important step towards the establishment of new protocols that support the ligation of remote DSB ends to achieve even larger rearrangements.
AU  - Brandl, C.
AU  - Ortiz, O.
AU  - Röttig, B.
AU  - Wefers, B.
AU  - Wurst, W.
AU  - Kühn, R.
C1  - 43005
C2  - 35950
SP  - 26-35
TI  - Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one-cell mouse embryos.
JO  - FEBS Open Bio
VL  - 5
PY  - 2015
SN  - 2211-5463
ER  -