کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8320631 | 1539399 | 2015 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Comparison of the kinetic parameters of the truncated catalytic subunit and holoenzyme of human DNA polymerase É
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کلمات کلیدی
dNTPPolδPCNADNA polymerase epsilonRPAssDNARFCPAGEdsDNADTTBSA - BSASingle-stranded DNA - DNA تک رشته ایdouble-stranded DNA - DNA رشته ایDNA polymerase α - DNA پلیمراز aDNA polymerase δ - DNA پلیمراز δbovine serum albumin - آلبومین سرم گاوProliferating Cell Nuclear Antigen - آنتیژن هسته ای تکثیر سلولیEDTA - اتیلن دی آمین تترا استیک اسید Ethylenediaminetetraacetic acid - اتیلینیدامین تتراستیک اسیدpolyacrylamide gel electrophoresis - الکتروفورز ژل پلی آکریل آمیدdithiothreitol - دیتیوتریتولPre-steady-state kinetics - سینتیک حالت پیش فرضReplication factor C - عامل تکرار Cpol - پلPolymerase - پلیمراز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
زیست شیمی
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Comparison of the kinetic parameters of the truncated catalytic subunit and holoenzyme of human DNA polymerase É Comparison of the kinetic parameters of the truncated catalytic subunit and holoenzyme of human DNA polymerase É](/preview/png/8320631.png)
چکیده انگلیسی
Numerous genetic studies have provided compelling evidence to establish DNA polymerase É (PolÉ) as the primary DNA polymerase responsible for leading strand synthesis during eukaryotic nuclear genome replication. PolÉ is a heterotetramer consisting of a large catalytic subunit that contains the conserved polymerase core domain as well as a 3â²Â â 5â² exonuclease domain common to many replicative polymerases. In addition, PolÉ possesses three small subunits that lack a known catalytic activity but associate with components involved in a variety of DNA replication and maintenance processes. Previous enzymatic characterization of the PolÉ heterotetramer from budding yeast suggested that the small subunits slightly enhance DNA synthesis by PolÉ in vitro. However, similar studies of the human PolÉ heterotetramer (hPolÉ) have been limited by the difficulty of obtaining hPolÉ in quantities suitable for thorough investigation of its catalytic activity. Utilization of a baculovirus expression system for overexpression and purification of hPolÉ from insect host cells has allowed for isolation of greater amounts of active hPolÉ, thus enabling a more detailed kinetic comparison between hPolÉ and an active N-terminal fragment of the hPolÉ catalytic subunit (p261N), which is readily overexpressed in Escherichia coli. Here, we report the first pre-steady-state studies of fully-assembled hPolÉ. We observe that the small subunits increase DNA binding by hPolÉ relative to p261N, but do not increase processivity during DNA synthesis on a single-stranded M13 template. Interestingly, the 3â²Â â 5â² exonuclease activity of hPolÉ is reduced relative to p261N on matched and mismatched DNA substrates, indicating that the presence of the small subunits may regulate the proofreading activity of hPolÉ and sway hPolÉ toward DNA synthesis rather than proofreading.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: DNA Repair - Volume 29, May 2015, Pages 16-22
Journal: DNA Repair - Volume 29, May 2015, Pages 16-22
نویسندگان
Walter J. Zahurancik, Andrey G. Baranovskiy, Tahir H. Tahirov, Zucai Suo,