Alexey Olovnikov: theoretical biology beyond the margins.

Alexey Olovnikov (1936-2022) is an author of the famous marginotomy hypothesis, where he recognized the DNA end replication problem and its role in cell aging. In this biographical note we celebrate the 50th anniversary of this theoretical discovery that later enjoyed a brilliant confirmation and gave rise to a new thriving field of molecular biology and gerontology. We also take a look at the evolution of ideas in Alexey Olovnikov's lifelong quest to uncover the molecular mechanisms of aging, exploring the reasons why he walked away from his initial conclusion about the key role of telomeres in aging, and built a new vast theoretical landscape that linked all stages of ontogenesis.

Telomeres in health and longevity: special issue in memory of Alexey Olovnikov.

In this special issue we commemorate theoretical biologist Alexey Olovnikov (1936-2022), whose theory of marginotomy has laid the foundation for the new field of biology that studies the molecular structure of telomeres and its role in health, longevity and aging. This issue contains a collection of reviews and research articles that discuss different aspects of telomere and telomerase research, ranging from telomere length dynamics in wild animal populations to problems of telomere maintenance during human space flight.

"If I Were in Nature's Place, I Would Do It Like This..." Life and Hypotheses of Alexey Olovnikov.

In this article, we commemorate the life and scientific journey of the brilliant gerontologist-theorist Alexey Olovnikov (1936-2022). In 1971, he published his famous "marginotomy" hypothesis, in which he predicted the replicative shortening of telomeres and its role as a counter of cell divisions and biological age of an organism. This work put forth several remarkable assumptions, including the existence of telomerase, which were confirmed two decades later. Despite this, Alexey Olovnikov moved further in his theoretical studies of aging and proposed a series of new hypotheses that seem no less exotic than the marginotomy hypothesis once appeared. Alexey Olovnikov had an extraordinary way of looking at biological problems and, in addition to aging, authored striking concepts about development, biorhythms, and evolution.

Homeostatic functions of the p53 tumor suppressor: regulation of energy metabolism and antioxidant defense.

The p53 tumor suppressor plays pivotal role in the organism by supervising strict compliance of individual cells to needs of the whole organisms. It has been widely accepted that p53 acts in response to stresses and abnormalities in cell physiology by mobilizing the repair processes or by removing the diseased cells through initiating the cell death programs. Recent studies, however, indicate that even under normal physiological conditions certain activities of p53 participate in homeostatic regulation of metabolic processes and that these activities are important for prevention of cancer. These novel functions of p53 help to align metabolic processes with the proliferation and energy status, to maintain optimal mode of glucose metabolism and to boost the energy efficient mitochondrial respiration in response to ATP deficiency. Additional activities of p53 in non-stressed cells tune up the antioxidant defense mechanisms reducing the probability of mutations caused by DNA oxidation under conditions of daily stresses. The deficiency in the p53-mediated regulation of glycolysis and mitochondrial respiration greatly accounts for the deficient respiration of the predominance of aerobic glycolysis in cancer cells (the Warburg effect), while the deficiency in the p53-modulated antioxidant defense mechanisms contributes to mutagenesis and additionally boosts the carcinogenesis process.

Efficient translation initiation directed by the 900-nucleotide-long and GC-rich 5' untranslated region of the human retrotransposon LINE-1 mRNA is strictly cap dependent rather than internal ribosome entry site mediated.

Retrotransposon L1 is a mobile genetic element of the LINE family that is extremely widespread in the mammalian genome. It encodes a dicistronic mRNA, which is exceptionally rare among eukaryotic cellular mRNAs. The extremely long and GC-rich L1 5' untranslated region (5'UTR) directs synthesis of numerous copies of RNA-binding protein ORF1p per mRNA. One could suggest that the 5'UTR of L1 mRNA contained a powerful internal ribosome entry site (IRES) element. Using transfection of cultured cells with the polyadenylated monocistronic (L1 5'UTR-Fluc) or bicistronic (Rluc-L1 5'UTR-Fluc) RNA constructs, capped or uncapped, it has been firmly established that the 5'UTR of L1 does not contain an IRES. Uncapping reduces the initiation activity of the L1 5'UTR to that of background. Moreover, the translation is inhibited by upstream AUG codons in the 5'UTR. Nevertheless, this cap-dependent initiation activity of the L1 5'UTR was unexpectedly high and resembles that of the beta-actin 5'UTR (84 nucleotides long). Strikingly, the deletion of up to 80% of the nucleotide sequence of the L1 5'UTR, with most of its stem loops, does not significantly change its translation initiation efficiency. These data can modify current ideas on mechanisms used by 40S ribosomal subunits to cope with complex 5'UTRs and call into question the conception that every long GC-rich 5'UTR working with a high efficiency has to contain an IRES. Our data also demonstrate that the ORF2 translation initiation is not directed by internal initiation, either. It is very inefficient and presumably based on a reinitiation event.

Sense transcripts originated from an internal part of the human retrotransposon LINE-1 5' UTR.

L1 (LINE-1) is one of the most abundant families of human transposable elements. Full-length human L1 has an ~900 bp long 5' untranslated region (5' UTR) which harbors an internal promoter for the RNA polymerase II. It is generally accepted that the first 100 bp of the 5' UTR function as a "minimal promoter" which directs transcription of the entire LINE-1 unit from the extreme 5' terminus. We re-investigated promoter activities of the different DNA fragments that cover the whole L1 5' UTR in cultured human cells by using the luciferase reporter system. Analysis of both mRNA expression and luciferase activity levels indicated that the very important region for the effective transcription is located within the internal part of the L1 5' UTR between nucleotide positions +390 and +526. 5' RACE analysis revealed that in the context of the complete 5' UTR, this part drives mRNA synthesis both from the canonical 5'-terminal transcription start site (TSS) and from within the internal region. In the absence of the first 100 bp, the L1 5' UTR efficiently directed transcription from aberrant TSSs located within its 3' proximal part or the ORF1. Finally, we analyzed transcripts originated from endogenous (genomic) L1 elements and identified two novel TSSs located at positions +525 and +570. We propose a model in which the internal part (390-526) of the L1 5' UTR plays a key role for recruitment of transcription initiation complex, which then may be either positioned onto the 5' terminally located "minimal promoter", or used proximately to direct 5' truncated RNA copy. Intriguingly, this internal regulatory element substantially overlaps with the region of the L1 5' UTR that is known to drive transcription in the opposite direction suggesting the existence of a common core for the bidirectional transcription.