NlaIII

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NlaIII is a type II restriction enzyme isolated from Neisseria lactamica . [1] As part of the restriction modification system, NlaIII is able to prevent foreign DNA from integrating into the host genome by cutting double stranded DNA into fragments at specific sequences. [2] This results in further degradation of the fragmented foreign DNA and prevents it from infecting the host genome. [3]

Contents

Recognition site of NlaIII with a red line indicating the cutting pattern NlaIII recognition site.png
Recognition site of NlaIII with a red line indicating the cutting pattern

NlaIII recognizes the palindromic and complementary DNA sequence of CATG/GTAC and cuts outside of the G-C base pairs. This cutting pattern results in sticky ends with GTAC overhangs at the 3' end. [4]

Characteristics

NlaIII from N. lactamica contains two key components: a methylase and an endonuclease. [5] The methylase is critical to recognition, while the endonuclease is used for cutting. [5] The gene (NlaIIIR) is 693 bp long and creates the specific 5’-CATG-3’ endonuclease. [6] A homolog of NlaIIIR is iceA1 from Helicobacter pylori . [6] In H. pylori, there exists a similar methylase gene called hpyIM which is downstream of iceA1. [7] ICEA1 is an endonuclease that also recognizes the 5’-CATG-3’ sequence. [6] IceA1 in H. pylori is similar to that of NlaIII in N. lactamica.

NlaIII contains an ICEA protein that encompasses the 4 to 225 amino acid region. [6] [8] H. pylori also contains the same protein. [9] H. pylori infection often leads to gastrointestinal issues such as peptic ulcers, gastric adenocarcinoma and lymphoma. [10] Researchers speculate that ICEA proteins serve as potential markers for gastric cancer [7]

Isoschizomers

NlaIII isoschizomers recognize and cut the same recognition sequence 5’-CATG-3’. [11] Endonucleases that cut at this sequence include:

Applications

NlaIII can be used in many different experimental procedures [12] such as:

References

  1. "nlaIIIR - Type-2 restriction enzyme NlaIII - Neisseria lactamica - nlaIIIR gene & protein". www.uniprot.org. Retrieved 2020-11-05.
  2. Sitaraman, Ramakrishnan (2016). "The Role of DNA Restriction-Modification Systems in the Biology of Bacillus anthracis". Frontiers in Microbiology. 7: 11. doi: 10.3389/fmicb.2016.00011 . ISSN   1664-302X. PMC   4722110 . PMID   26834729.
  3. Berg, Jeremy M.; Tymoczko, John L.; Stryer, Lubert (2002). "Restriction Enzymes: Performing Highly Specific DNA-Cleavage Reactions". Biochemistry. 5th Edition.
  4. 1 2 Pasternak, Jack J. (2005). An introduction to human molecular genetics: mechanisms of inherited diseases (2nd ed.). Hoboken, N.J: John Wiley & Sons. ISBN   978-0-471-71917-5.
  5. 1 2 Morgan, R. D.; Camp, R. R.; Wilson, G. G.; Xu, S. Y. (1996-12-12). "Molecular cloning and expression of NlaIII restriction-modification system in E. coli". Gene. 183 (1–2): 215–218. doi:10.1016/s0378-1119(96)00561-6. ISSN   0378-1119. PMID   8996109.
  6. 1 2 3 4 Xu, Qing; Morgan, R. D.; Roberts, R. J.; Xu, S. Y.; van Doorn, L. J.; Donahue, J. P.; Miller, G. G.; Blaser, Martin J. (2002-09-01). "Functional analysis of iceA1, a CATG-recognizing restriction endonuclease gene in Helicobacter pylori". Nucleic Acids Research. 30 (17): 3839–3847. doi: 10.1093/nar/gkf504 . ISSN   0305-1048. PMC   137426 . PMID   12202769.
  7. 1 2 Xu, Q.; Peek, R. M.; Miller, G. G.; Blaser, M. J. (1997-11-01). "The Helicobacter pylori genome is modified at CATG by the product of hpyIM". Journal of Bacteriology. 179 (21): 6807–6815. doi: 10.1128/jb.179.21.6807-6815.1997 . ISSN   0021-9193. PMC   179612 . PMID   9352933.
  8. "UniProtKB - Q51083 (T2N3_NEILA)". UniProt. Retrieved December 2, 2020.
  9. Wong, Benjamin Chun Yu; Yin, Yan; Berg, Douglas E.; Xia, Harry Hua-Xiang; Zhang, Jian Zhong; Wang, Wei Hong; Wong, Wai Man; Huang, Xiao Ru; Tang, Vera Shun Yim; Lam, Shiu Kum (2001). "Distribution of Distinct vacA, cagA and iceA Alleles in Helicobacter pylori in Hong Kong" . Helicobacter. 6 (4): 317–324. doi:10.1046/j.1523-5378.2001.00040.x. ISSN   1523-5378. PMID   11843964. S2CID   10084164.
  10. Kusters, Johannes G.; Vliet, Arnoud H. M. van; Kuipers, Ernst J. (2006-07-01). "Pathogenesis of Helicobacter pylori Infection". Clinical Microbiology Reviews. 19 (3): 449–490. doi: 10.1128/CMR.00054-05 . ISSN   0893-8512. PMC   1539101 . PMID   16847081.
  11. "Enzyme Finder". New England Biolabs. Retrieved 23 October 2020.
  12. "Hin1II (NlaIII) (5 U/µL)". Thermo Fisher Scientific. Retrieved November 3, 2020.