Pradhan, S., Kalia, I., Roy, S. S., Singh, O. P., Adak, T., Singh, A. P. and Dhar, S. K. (2019) Molecular characterization and expression profile of an alternate proliferating cell nuclear antigen homolog PbPCNA2 in Plasmodium berghei. IUBMB Life, 71(9), pp. 1293-1301. (doi: 10.1002/iub.2036) (PMID:30865364)
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Abstract
Proliferative cell nuclear antigen (PCNA) is the processivity factor for various DNA polymerases and it functions in response to DNA damage in eukaryotic system. Plasmodium falciparum contains two PCNAs, while PCNA1 has been attributed to DNA replication, the role of PCNA2 has been assigned to DNA damage response in erythrocytic developmental stages. Although a recent transposon mediated knockout strategy qualified pcna2 as a nonessential gene in Plasmodium berghei, a conventional homologous recombination-based knockout strategy has not been employed for this gene yet. Moreover, the cellular dynamics of PCNA2 in extraerythrocytic stages still remain elusive in Plasmodium. We attempted multiple times to knock out PbPCNA2 from the parasite genome using homologous recombination strategy without much success. However, we were able to generate PbPCNA2-GFP tagged transgenic parasites confirming that the pcna2 locus is amenable to genetic manipulation. The GFP-tagged parasites showed similar growth phenotype, compared to wild-type parasites, in both erythrocytic and sporogonic cycle, suggesting that tagging had no effect on parasite physiology. PbPCNA2 expression was also observed during the sporogonic cycle in midgut oocyst and salivary gland sporozoites. The PbPCNA2 expression was upregulated in the presence of DNA damaging agents like hydroxyurea and methyl methanesulphonate. Our inability to knock out PCNA2 suggested its essentiality in the parasite development and elevated expression during DNA damaging condition hint at a critical role of the protein in parasite physiology.
| Item Type: | Articles |
|---|---|
| Additional Information: | This work is supported by the grants from Department of Biotechnology, Govt. of India (BT/PR15639/Med/29/1145/2016 to SKD, BT/PR21569/NNT/28/1234/2017 jointly to SKD and APS). Financial support in part also came from the SERB, DST Govt. of India grant (EMR/2015/001546) to APS. SKD also acknowledges UGC-SAP, DST-PURSE, and UPE-II funding (ID no. 28) awarded to JNU for financial support. SP acknowledges UGC, India, and SSR acknowledge UGC (DS Kothari Postdoctoral Fellowship), India, for fellowships. Fellowship to IK was supported by NII core grants. |
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Pradhan, Dr Sabyasachi |
| Authors: | Pradhan, S., Kalia, I., Roy, S. S., Singh, O. P., Adak, T., Singh, A. P., and Dhar, S. K. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine |
| Journal Name: | IUBMB Life |
| ISSN: | 1521-6543 |
| ISSN (Online): | 1521-6551 |
| Published Online: | 13 March 2019 |
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