Telomerase paper课件.pptVIP

Extension of Life-Span by Introduction of Telomerase into Normal Human Cells. Bodner et. al. Science 279, 349-352 (1998) From the laboratory of Woodring E. Wright This paper is an important paper because it shows direct causal relationship between telomerase expression and cellular life-span. Normal human diploid cells have a finite proliferative life-span when placed in culture. After this, they enter a non-proliferative state termed senescence. Replicative senescence is dependent on cumulative cell divisions and not chronologic or metabolic time. Proliferation is limited by a “mitotic clock”. Replicative senescence was noted over 30 years ago by Hayflick. (Hayflick limit) Telomere loss is thought to control the entry into senescent state. Human telomeres consist of the sequence TTAGGG and these repeats are synthesized by a ribonucleoprotein enzyme called telomerase. Telomerase is active in germline cells and in humans the telomeres are maintained in germline cells at about 15 kbp length. In contrast, telomerase is not expressed in somatic cells and telomere length is significantly shorter in somatic tissues. The telomere hypothesis of aging proposes that cells become senescent when progressive telomere shortening during each cell division produces a threshold telomere length. Human telomerase reverse transcriptase subunit: hTRT. Telomerase activity in normal human diploid cells can be reconstituted by expressing hTRT in normal diploid cells, which express low levels of the template RNA component of the enzyme but do not express hTRT. This provides a method to manipulate telomere length and directly test the hypothesis that telomere shortening causes cellular senecscence. In this paper, the authors have reconstituted the telomerase activity in human cells, thereby lengthening their telomeres and asked the question as to how this would affect the life span and/or senescence of these cells. Three different primary cells in culture were used. Retinal Pigm