The large subunit of Leishmania topoisomerase I functions as the 'molecular steer' in type IB topoisomerase.

Abstract:

:Kinetoplastid topoisomerase IB is an unusual bisubunit enzyme where reconstitution of the large (LdTOPIL or L) and small (LdTOPIS or S) subunits shows functional activity. It is yet to be deciphered whether one subunit or both navigate the heterodimer to its cellular DNA targets. Tethering a specific DNA-binding protein to topoisomerase I alters its site specificity. The chimeric constructs UMSBP-LdTOPIL/S or U-L/S (fusion of UMSBP to the N-terminus of L and reconstituted with S) and LdTOPIL/UMSBP-LdTOPIS or L/U-S (fusion of UMSBP to the N-terminus of S and reconstituted with L) exhibit relaxation activity. Only U-L/S shows altered site specificity and enhanced DNA-binding affinity for the universal minicircle sequence (UMS) containing substrate. This proves that L alone serves as the 'molecular steer' for this heterodimer. Reconstituted U-L/S also induces cleavage close to UMS and causes minicircle linearization. The differential properties of the reconstituted chimeras U-L/S and L/U-S reveal the structural and functional asymmetry between the heterodimer. Therefore this study helps in a better understanding of the mechanistic details underlying topoisomerization by this bi-subunit enzyme.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

BoseDasgupta S,Ganguly A,Das BB,Roy A,Khalkho NV,Majumder HK

doi

10.1111/j.1365-2958.2007.06002.x

subject

Has Abstract

pub_date

2008-01-01 00:00:00

pages

31-46

issue

1

eissn

0950-382X

issn

1365-2958

pii

MMI6002

journal_volume

67

pub_type

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