4. Kaplan M et al (2016) EGFR dynamics change
during activation in native membranes as
revealed by NMR. Cell 167:1241–1251.e11
5. Oikonomou CM, Jensen GJ (2017) A new
view into prokaryotic cell biology from electron cryotomography. Nat Rev Microbiol
15:128
6. Luc ˇic ´ V, Rigort A, Baumeister W (2013) Cryoelectron tomography: the challenge of doing
structural biology in situ. J Cell Biol
202:407–419
7. Chang Y-W et al (2016) Architecture of the
type IVa pilus machine. Science 351:aad2001
8. Kaplan M et al (2019) The presence and
absence of periplasmic rings in bacterial flagellar motors correlates with stator type. elife 8:
e43487
9. Chaban B, Coleman I, Beeby M (2018) Evolution of higher torque in campylobacter-type
bacterial flagellar motors. Sci Rep 8:97
10. Kaplan M et al (2019) In situ imaging of the
bacterial flagellar motor disassembly and
assembly processes. EMBO J 38:e100957.
https://doi.org/10.15252/embj.
2018100957
11. Ferreira JL et al (2019) γ-proteobacteria eject
their polar flagella under nutrient depletion,
retaining flagellar motor relic structures. PLoS
Biol 17:e3000165. https://doi.org/10.1101/
367458
12. Zhu S et al (2019) In situ structures of polar
and lateral flagella revealed by Cryo-Electron
tomography. J Bacteriol 201:13
13. Zhao X et al (2013) Cryoelectron tomography
reveals the sequential assembly of bacterial flagella in Borrelia burgdorferi. Proc Natl Acad
Sci 110:14390–14395
14. Hagen WJH, Wan W, Briggs JAG (2017)
Implementation of a cryo-electron tomography tilt-scheme optimized for high resolution
subtomogram averaging. J Struct Biol
197:191–198
15. Briggs JAG (2013) Structural biology in situ—
the potential of subtomogram averaging. Curr
Opin Struct Biol 23:261–267
16. Leigh KE et al (2019) Subtomogram averaging
from cryo-electron tomograms. Methods Cell
Biol 152:217–259
17. Murphy GE, Leadbetter JR, Jensen GJ (2006)
In situ structure of the complete Treponema
primitia
flagellar
motor.
Nature
442:1062–1064
18. Chen S et al (2011) Structural diversity of bacterial flagellar motors: structural diversity of
bacterial
flagellar
motors.
EMBO
J
30:2972–2981
19. Zhao X, Norris SJ, Liu J (2014) Molecular
architecture of the bacterial flagellar motor in
cells. Biochemistry 53:4323–4333
20. Gold VA, Salzer R, Averhoff B, Ku ¨hlbrandt W
(2015) Structure of a type IV pilus machinery
in the open and closed state. elife 4:e07380
21. Hu B, Lara-Tejero M, Kong Q, Gala ´n JE, Liu J
(2017) In situ molecular architecture of the
salmonella type III secretion machine. Cell
168:1065–1074.e10
22. Ghosal D, Chang Y-W, Jeong KC, Vogel JP,
Jensen GJ (2017) In situ structure of the legionella dot/Icm type IV secretion system by electron
cryotomography.
EMBO
Rep
18:726–732
23. Chang Y-W, Shaffer CL, Rettberg LA,
Ghosal D, Jensen GJ (2018) In vivo structures
of the helicobacter pylori cag type IV secretion
system. Cell Rep 23:673–681
24. Ghosal D et al (2019) Molecular architecture,
polar targeting and biogenesis of the legionella
Dot/Icm T4SS. Nat Microbiol 4:1173–1182
25. Yang W et al (2019) In situ conformational
changes of the Escherichia coli serine chemoreceptor in different signaling states. MBio 10:
e00973-19
26. Rapisarda C et al (2019) In situ and highresolution cryo-EM structure of a bacterial
type VI secretion system membrane complex.
EMBO J 38:e100886
27. Wan W et al (2017) Structure and assembly of
the Ebola virus nucleocapsid. Nature
551:394–397
28. Schur FKM et al (2016) An atomic model of
HIV-1 capsid-SP1 reveals structures regulating
assembly
and
maturation.
Science
353:506–508
29. Rigort A et al (2012) Focused ion beam micromachining of eukaryotic cells for cryoelectron
tomography.
Proc
Natl
Acad
Sci
109:4449–4454
30. Danev R, Buijsse B, Khoshouei M, Plitzko JM,
Baumeister W (2014) Volta potential phase
plate for in-focus phase contrast transmission
electron microscopy. Proc Natl Acad Sci U S A
111:15635–15640
31. Chreifi G, Chen S, Metskas LA, Kaplan M,
Jensen GJ (2019) Rapid tilt-series acquisition
for electron cryotomography. J Struct Biol
205:163–169
32. Ghosal D, Kaplan M, Chang Y-W, Jensen GJ
(2019) In situ imaging and structure determination of bacterial toxin delivery systems using
electron cryotomography. In: Buchrieser C,
Hilbi H (eds) Legionella, vol 1921. Springer,
New York, pp 249–265
110
Mohammed Kaplan et al.
during activation in native membranes as
revealed by NMR. Cell 167:1241–1251.e11
5. Oikonomou CM, Jensen GJ (2017) A new
view into prokaryotic cell biology from electron cryotomography. Nat Rev Microbiol
15:128
6. Luc ˇic ´ V, Rigort A, Baumeister W (2013) Cryoelectron tomography: the challenge of doing
structural biology in situ. J Cell Biol
202:407–419
7. Chang Y-W et al (2016) Architecture of the
type IVa pilus machine. Science 351:aad2001
8. Kaplan M et al (2019) The presence and
absence of periplasmic rings in bacterial flagellar motors correlates with stator type. elife 8:
e43487
9. Chaban B, Coleman I, Beeby M (2018) Evolution of higher torque in campylobacter-type
bacterial flagellar motors. Sci Rep 8:97
10. Kaplan M et al (2019) In situ imaging of the
bacterial flagellar motor disassembly and
assembly processes. EMBO J 38:e100957.
https://doi.org/10.15252/embj.
2018100957
11. Ferreira JL et al (2019) γ-proteobacteria eject
their polar flagella under nutrient depletion,
retaining flagellar motor relic structures. PLoS
Biol 17:e3000165. https://doi.org/10.1101/
367458
12. Zhu S et al (2019) In situ structures of polar
and lateral flagella revealed by Cryo-Electron
tomography. J Bacteriol 201:13
13. Zhao X et al (2013) Cryoelectron tomography
reveals the sequential assembly of bacterial flagella in Borrelia burgdorferi. Proc Natl Acad
Sci 110:14390–14395
14. Hagen WJH, Wan W, Briggs JAG (2017)
Implementation of a cryo-electron tomography tilt-scheme optimized for high resolution
subtomogram averaging. J Struct Biol
197:191–198
15. Briggs JAG (2013) Structural biology in situ—
the potential of subtomogram averaging. Curr
Opin Struct Biol 23:261–267
16. Leigh KE et al (2019) Subtomogram averaging
from cryo-electron tomograms. Methods Cell
Biol 152:217–259
17. Murphy GE, Leadbetter JR, Jensen GJ (2006)
In situ structure of the complete Treponema
primitia
flagellar
motor.
Nature
442:1062–1064
18. Chen S et al (2011) Structural diversity of bacterial flagellar motors: structural diversity of
bacterial
flagellar
motors.
EMBO
J
30:2972–2981
19. Zhao X, Norris SJ, Liu J (2014) Molecular
architecture of the bacterial flagellar motor in
cells. Biochemistry 53:4323–4333
20. Gold VA, Salzer R, Averhoff B, Ku ¨hlbrandt W
(2015) Structure of a type IV pilus machinery
in the open and closed state. elife 4:e07380
21. Hu B, Lara-Tejero M, Kong Q, Gala ´n JE, Liu J
(2017) In situ molecular architecture of the
salmonella type III secretion machine. Cell
168:1065–1074.e10
22. Ghosal D, Chang Y-W, Jeong KC, Vogel JP,
Jensen GJ (2017) In situ structure of the legionella dot/Icm type IV secretion system by electron
cryotomography.
EMBO
Rep
18:726–732
23. Chang Y-W, Shaffer CL, Rettberg LA,
Ghosal D, Jensen GJ (2018) In vivo structures
of the helicobacter pylori cag type IV secretion
system. Cell Rep 23:673–681
24. Ghosal D et al (2019) Molecular architecture,
polar targeting and biogenesis of the legionella
Dot/Icm T4SS. Nat Microbiol 4:1173–1182
25. Yang W et al (2019) In situ conformational
changes of the Escherichia coli serine chemoreceptor in different signaling states. MBio 10:
e00973-19
26. Rapisarda C et al (2019) In situ and highresolution cryo-EM structure of a bacterial
type VI secretion system membrane complex.
EMBO J 38:e100886
27. Wan W et al (2017) Structure and assembly of
the Ebola virus nucleocapsid. Nature
551:394–397
28. Schur FKM et al (2016) An atomic model of
HIV-1 capsid-SP1 reveals structures regulating
assembly
and
maturation.
Science
353:506–508
29. Rigort A et al (2012) Focused ion beam micromachining of eukaryotic cells for cryoelectron
tomography.
Proc
Natl
Acad
Sci
109:4449–4454
30. Danev R, Buijsse B, Khoshouei M, Plitzko JM,
Baumeister W (2014) Volta potential phase
plate for in-focus phase contrast transmission
electron microscopy. Proc Natl Acad Sci U S A
111:15635–15640
31. Chreifi G, Chen S, Metskas LA, Kaplan M,
Jensen GJ (2019) Rapid tilt-series acquisition
for electron cryotomography. J Struct Biol
205:163–169
32. Ghosal D, Kaplan M, Chang Y-W, Jensen GJ
(2019) In situ imaging and structure determination of bacterial toxin delivery systems using
electron cryotomography. In: Buchrieser C,
Hilbi H (eds) Legionella, vol 1921. Springer,
New York, pp 249–265
110
Mohammed Kaplan et al.