Anthony V. Furano, M.D.
Genomic Structure and Function Section, Laboratory of Cell and Molecular Biology
Building 8, Room 201
8 Center Dr
Bethesda, MD 20814
+1 301 496 6180
(1) Understanding replication of the L1 retrotransposon, an intragenomic parasite that has generated at least 50% of human DNA and is still active in modern humans. (2) Understanding the derangements in DNA repair that can induce mutations in normal flanking DNA.
Mammalian L1 elements are obligate intragenomic parasites that replicate (retrotranspose) by copying their RNA transcripts (and those of other genes) into genomic DNA, a reaction that is mediated by two L1 proteins, ORF1p and ORF2p, which are regulated by the L1 5’ untranslated region (UTR). L1 has been amplifying and evolving in mammalian lineages for approximately the last 100 million years. During this time at least 16 successive L1 families arose, only to have gone extinct and then be supplanted by a novel active family. Having generated at least 50% of human DNA, L1 activity has had a defining effect on the evolution, structure, and function of the human genomes. Despite being seriously deleterious and a target of numerous host repressive mechanisms, L1 activity and evolution is ongoing in modern humans, where it can damage DNA and cause genetic defects. It can also be re-activated in somatic cells including tumors, neuronal progenitor cells, and consequent to aging. Not only can L1 insertions cause genetic defects, but, as ORF1p activity depends on phosphorylation of its conserved protein kinase target sites, it can compete for or perturb the critical phosphokinase cascades. Given its generally deleterious, and sometimes catastrophic effects accounting for the persistence of L1 over evolutionary time is a major issue for mammalian biology. Foundational, pre-whole genome studies showed that evolutionary change in L1 was often associated with the emergence of novel L1 families. These included the emergence of novel 5’UTRs or recurrent instances of remodeling of the ORF1p coiled coil. Such evolutionary novelty was presumably related to maintain the biochemical competence of L1, which is the ultimate arbiter of evolution. Additionally, the DNA inserts of the extinct L1 families remain in the genome where they accumulate mutations at the neutral mutation rate. They thereby represent an unparalleled source of DNA fossils that provide a genetic record of its host, our analysis of which prompted our hypothesis the DNA repair can be mutagenic, which we demonstrated experimentally and which now constitutes one of our major pursuits.
Applying our Research
As mammals have coevolved with the L1 parasite, understanding the biochemistry of L1 replication is essential to fully understand the biology of mammals, and assessing the potential, and perhaps existential, effect of L1 on current and future human populations. More particularly, in addition to having already massively altered the structure of mammalian genomes, L1 activity in present human populations is sufficiently active to have reduced their genetic fitness. While de novo in utero L1 insertions are an important cause of genetic defects, ongoing L1 activity not only constitutes a nonending assault on the genetic integrity of human genomes, but it also continually generates evolutionary novel L1 elements that could wreak havoc on human populations if they can evade host mechanisms that repress them. Such an event would be much like the emergence of a deadly pandemic-producing novel strain of influenza virus. In fact, the phylodynamics of L1 evolution and that of the HA epitope of influenza virus are uncannily similar, suggesting that similar dynamics of the host/parasite relationship have governed both.
Equally relevant to the genetic well-being of human populations is determining the factors that affect the neutral mutation rate (the rate at which base substitutions accumulate). Base substitutions are a major source of genetic variation and genetic diseases, and alterations of the base substitution rate accompany the progression of many cancers. Our suggestion that DNA repair itself could be mutagenic has profound implications for understanding the determinants of the neutral mutation rate, for the use of this rate in evolutionary studies, and for analyses that rely on sequence conservation, or lack thereof, as a proxy for natural selection. We have now supported this idea experimentally in vivo using a system wherein we can introduce defined DNA lesions in various sequence contexts and showed that their repair induces mutations in normal flanking DNA. This system constitutes an invaluable asset for the biochemical analysis of repair-induced mutagenesis, for analyzing the mutational propensity of different human cancers, and their implications for human disease and therapeutics.
- Deputy Laboratory Chief, LCMB, NIDDK, NIH, 2011-present
- Acting Laboratory Chief, LMCB, NIDDK, NIH, 2006-2011
- Medical Officer (Research), Section Chief, GSF, LMCB, NIDDK, NIH, 1983-2006
- Editorial board, Mobile DNA, 2009-present
- Lecturer on LINEs & SINEs: George Washington University, BIOC 254 Fundamentals of Molecular Biology, 2005-2010
- Contributing Member of the USCD Project for Explaoining the Origins of Humans (La Jolla project on the origin of humans, LOH), 2005-present
- B.S., Tufts University
- M.D., Yale University
Chen J, Miller BF, Furano AV. Repair of naturally occurring mismatches can induce mutations in flanking DNA. Elife. 2014;3:e02001.
Naufer MN, Callahan KE, Cook PR, Perez-Gonzalez CE, Williams MC, Furano AV. L1 retrotransposition requires rapid ORF1p oligomerization, a novel coiled coil-dependent property conserved despite extensive remodeling. Nucleic Acids Res. 2016;44(1):281-93.
Naufer MN, Furano AV, Williams MC. Protein-nucleic acid interactions of LINE-1 ORF1p. Semin Cell Dev Biol. 2018.
Chen J, Furano AV. Breaking bad: The mutagenic effect of DNA repair. DNA Repair (Amst). 2015;32:43-51.
Cook PR, Jones CE, Furano AV. Phosphorylation of ORF1p is required for L1 retrotransposition. Proc Natl Acad Sci U S A. 2015;112(14):4298-303.
Related Scientific Focus Areas
Genetics and Genomics
Molecular Biology and Biochemistry
This page was last updated on August 1st, 2019