bioRxiv Science⌕ Search

Biology subjects

Parkar, S. N.

Publications and source records attributed to Parkar, S. N..

2 recordsLinked to original sources

Δ246p53 is a new p53 isoform that responds to DNA damage and regulates tumour growth

p53 is with little doubt one of the most powerful genes in our genome, as it makes growth vs arrest, repair vs replacement, catabolism vs anabolism, life vs death decisions in the cell. An alteration or malfunction in p53 may lead to cancer, degeneration or premature ageing. So, it is not surprising that p53 is also one of the most complex and tightly regulated genes, encoding for at least 10 RNA variants and 12 widely accepted protein forms. Here we identify one new p53 protein isoform of approximately 18 kDa that we termed {Delta}246p53. {Delta}246p53 is translated from an alternative translation initiation site (TIS) in codon 246. TIS-246 is preceded by a strong Kozak sequence and appears conserved in vertebrates, from sea lamprey to humans. {Delta}246p53s origin and expression in cells were confirmed by frameshift and start codon mutations; five different antibodies against several epitopes from its N-terminus down to its C-terminus and one against the DNA binding domain region just upstream of its initiator methionine working as negative control; as well as siRNAs and an antisense oligo targeting TIS-246, which knocked-down {Delta}246p53 with little or no effect on full-length (FL) p53 protein levels. {Delta}246p53 was induced by DNA damage and triggered senescence and impaired tumour formation/growth in colony formation assays. Mechanistically, {Delta}246p53 interacted with FLp53, leading to a decrease in the expression of downstream target genes HDM2 and p21. Concurrently, it also interacted with {Delta}Np63, leading to p53-independent p63-mediated activation of p21, a known senescence activator. Our results unveil a new naturally occurring and tightly controlled factor with specific functions in p21 regulation and senescence. Future studies on {Delta}246p53 are likely to help us better understand and control the processes of tumour suppression and ageing.

cancer biology↗

Internal Translation of p53 Oncoproteins During Integrated Stress Response Confers Survival Advantage on Cancer Cells

p53 is the most known and studied tumour suppressor gene. Yet we have recently shown that p53 is also a proto-oncogene, as it encodes the {Delta}160p53 oncoprotein. Integrated stress response (ISR) is a survival pathway frequently activated in cancers, marked by the phosphorylation of eukaryotic initiation factor 2alpha (eIF2) and a defined reprogramming in mRNA translation. Here we identified ISR as a powerful trigger of p53 oncogene, leading to the induction of not only {Delta}160p53 but also {Delta}133p53, another protein variant of the p53 gene. Upon ISR the two isoforms were translated internally from p53 full-length (FL) transcript through an internal regulator of expression site (IRES) located in the vicinity of codon 160. Frameshift mutations upstream of codons 133 and 160 demonstrated that FLp53 protein synthesis is not required for making {Delta}133p53 and {Delta}160p53. Instead, targeting IRES(160) with an antisense oligo was sufficient to efficiently and specifically impair the expression of these isoforms without affecting FLp53 levels. This in turn averted ISRs protective program culminating in cancer cell cycle arrest and death. Mechanistically, FLp53 showed 3 times more affinity to {Delta}160p53 than to other isoforms, {Delta}40p53 or {Delta}133p53. During ISR {Delta}160p53 localized to the nucleus and strongly inhibited FLp53-mediated activation of pro-apoptotic gene p53 upregulated modulator of apoptosis (PUMA). Our results uncover a new branch of the ISR network essential for cancer cell survival and growth and establish the proof of concept for a new strategy to target cancer.

cancer biology↗